Kinase inhibitors

ABSTRACT

The invention relates to inhibitors of kinases, compositions comprising the inhibitors, and methods of using the inhibitors and inhibitor compositions. The inhibitors and compositions comprising them are useful for treating disease or disease symptoms. The invention also provides for methods of making kinase inhibitor compounds, methods of inhibiting kinase activity, and methods for treating disease or disease symptoms.

[0001] This application claims priority benefit under Title 35 USC§119(e) of U.S. Provisional Application No. 60/183,256 filed Feb. 17,2000 and entitled Kinase Inhibitors, the entire contents of which areincorporated herein by reference.

BACKGROUND OF THE INVENTION

[0002] The invention relates to inhibitors of enzymes that catalyzephosphoryl transfer and/or that bind ATP/GTP nucleotides, compositionscomprising the inhibitors, and methods of using the inhibitors andinhibitor compositions. The inhibitors and compositions comprising themare useful for treating or modulating disease in which phosphoryltransferases, including kinases, may be involved, symptoms of suchdisease, or the effect of other physiological events mediated byphosphoryl transferases, including kinases. The invention also providesfor methods of making the inhibitor compounds and methods for treatingdiseases in which one or more phosphoryl transferase, including kinase,activities is involved.

[0003] Phosphoryl transferases are a large family of enzymes thattransfer phosphorous-containing groups from one substrate to another. Bythe conventions set forth by the Nomenclature Committee of theInternational Union of Biochemistry and Molecular Biology (IUBMB)enzymes of this type have Enzyme Commission (EC) numbers starting with2.7.-.- (See, Bairoch A., The ENZYME database in Nucleic Acids Re.s28:304-305(2000)). Kinases are a class of enzymes that function in thecatalysis of phosphoryl transfer. The protein kinases constitute thelargest subfamily of structurally related phosphoryl transferases andare responsible for the control of a wide variety of signal transductionprocesses within the cell. (See, Hardie, G. and Hanks, S. (1995) TheProtein Kinase Facts Book, I and II, Academic Press, San Diego, Calif.).Protein kinases are thought to have evolved from a common ancestral genedue to the conservation of their structure and catalytic function.Almost all kinases contain a similar 250-300 amino acid catalyticdomain. The protein kinases may be categorized into families by thesubstrates they phosphorylate (e.g., protein-tyrosine,protein-serine/threonine, histidine, etc.). Protein kinase sequencemotifs have been identified that generally correspond to each of thesekinase families (See, for example, Hanks, S. K., Hunter, T., FASEB J.,9:576-596 (1995); Knighton et al., Science, 253:407-414 (1991); Hiles etal., Cell, 70:419-429 (1992); Kunz et al., Cell, 73:585-596 (1993);Garcia-Bustos et al., EMBO J., 13:2352-2361 (1994)). Lipid kinases (e.g.PI3K) constitute a separate group of kinases with structural similarityto protein kinases.

[0004] Since the X-ray structure of the catalytic subunit ofcAMP-dependent protein kinase (cAPK) was elucidated, approximately twodozen additional protein kinase structures and one lipid kinasestructure have been solved as either apo enzymes or binary and ternarycomplexes (with ATP, ATP analogs, metal ions, ADP, ATP competitiveinhibitors in the absence or presence of peptide substrate or peptideinhibitors). These proteins share structurally conserved catalyticdomains (kinase domains) comprising two lobes that can be furthersubdivided into twelve subdomains. The N-terminal portion forms thesmall lobe (including subdomains I-IV) whose architecture is composed ofan antiparallel five-strand β-sheet and one α-helix, while the lowerC-terminal domain forms another lobe (including subdomains VIA-XI)containing mostly α-helical architecture. Subdomain V spans the twolobes. The N-terminal domain is thought to participate in orienting thenucleotide (or other binding entity), while the C-terminal domain isthought to be responsible for binding peptide substrate and initiatingphosphotransfer to the hydroxyl group of a serine, threonine, ortyrosine residue.

[0005] The N- and C-terminal domains are connected through a singlepeptide strand, to which the adenine moiety of ATP and/or GTP binds viaan eleven membered hydrogen bond cycle, involving the N1 and the N6amino group, and the backbone carbonyl and NH functions of twononconsecutive residues. This linker acts as a hinge about which thedomains can rotate with respect to each other without disruption of thesecondary architecture of the kinase. Several torsion angle changes inthe linker backbone allow this movement to occur. The ribose group ofATP is anchored to the enzyme via hydrogen bonds with residues withinthe ribose-binding pocket. The triphosphate group is held in positionvia various polar interactions with several variable residues from theglycine rich loop, the conserved DFG motif and the catalytic loop.

[0006] The “kinase domain” appears in a number of polypeptides whichserve a variety of functions. Such polypeptides include, for example,transmembrane receptors, intracellular receptor associated polypeptides,cytoplasmic located polypeptides, nuclear located polypeptides andsubcellular located polypeptides. The activity of protein kinases can beregulated by a variety of mechanisms. It must be noted, however, that anindividual protein kinase may be regulated by more than one mechanism.These mechanisms include, for example, autophosphorylation,transphosphorylation by other kinases, protein-protein interactions,protein-lipid interactions, protein-polynucleotide interactions, ligandbinding, and post-translational modification.

[0007] Protein and lipid kinases regulate many different cell processesincluding, but not limited to, proliferation, growth, differentiation,metabolism, cell cycle events, apoptosis, motility, transcription,translation and other signaling processes, by adding phosphate groups totargets such as proteins or lipids. Phosphorylation events catalyzed bykinases act as molecular on/off switches that can modulate or regulatethe biological function of the target protein. Phosphorylation of targetproteins occurs in response to a variety of extracellular signals(hormones, neurotransmitters, growth and differentiation factors, etc.),cell cycle events, environmental or nutritional stresses, etc. Proteinand lipid kinases can function in signaling pathways to activate orinactivate, or modulate the activity of (either directly or indirectly)the targets. These targets may include, for example, metabolic enzymes,regulatory proteins, receptors, cytoskeletal proteins, ion channels orpumps, or transcription factors. Uncontrolled signaling due to defectivecontrol of protein phosphorylation has been implicated in a number ofdiseases and disease conditions, including, for example, inflammation,cancer, allergy/asthma, disease and conditions of the immune system,disease and conditions of the central nervous system (CNS),cardiovascular disease, dermatology, and angiogenesis.

[0008] Initial interest in protein kinases as pharmacological targetswas stimulated by the findings that many viral oncogenes encodestructurally modified cellular protein kinases with constitutive enzymeactivity. These findings pointed to the potential involvement ofoncogene related protein kinases in human proliferative disorders.Subsequently, deregulated protein kinase activity, resulting from avariety of more subtle mechanisms, has been implicated in thepathophysiology of a number of important human disorders including, forexample, cancer, CNS conditions, and immunologically related diseases.The development of selective protein kinase inhibitors that can blockthe disease pathologies and/or symptoms resulting from aberrant proteinkinase activity has therefore generated much interest.

SUMMARY OF THE INVENTION

[0009] The invention relates to compounds of the formula:

[0010] wherein,

[0011] Each R¹ and R² is independently R³; R⁸; NHR³; NHR⁵; NHR⁶; NR⁵R⁵;NR⁵R⁶; SR⁵; SR⁶; OR⁵; OR⁶; C(O)R³; heterocyclyl optionally substitutedwith 1-4 independent R⁴ on each ring; or C1-C10 alkyl substituted with1-4 independent R⁴;

[0012] Each R³ is independently aryl; phenyl optionally substituted with1-4 independent R⁴; or heteroaryl optionally substituted with 1-4independent R⁴ on each ring; and the remaining variables are as definedherein.

[0013] The invention also relates to compositions comprising thesecompounds, methods of making these compounds, methods of inhibitingenzyme activity, particularly kinase activity, through use of thesecompounds, and methods of treating disease or disease symptoms in amammal, particularly where modulation of enzyme activity, and moreparticularly kinase activity, can affect disease outcome.

DETAILED DESCRIPTION OF THE INVENTION

[0014] The invention provides compounds useful in inhibiting kinaseactivity and inhibiting kinases or other polypeptides having sequencesor subsequences homologous to kinase sequences or subsequences. In oneembodiment, the inhibitory compound has the formula:

[0015] wherein,

[0016] Each R¹ and R² is independently R³; R⁸; NHR³; NHR⁵; NHR⁶; NR⁵R⁵;NR⁵R⁶; SR⁵; SR⁶; OR⁵; OR⁶; C(O)R³; heterocyclyl optionally substitutedwith 1-4 independent R⁴ on each ring; or C1-C10 alkyl substituted with1-4 independent R⁴;

[0017] Each R³ is independently aryl; phenyl optionally substituted with1-4 independent R⁴; or heteroaryl optionally substituted with 1-4independent R⁴ on each ring;

[0018] Each m is independently 0, 1, 2 or 3;

[0019] Each n is independently 1 or 2;

[0020] Each X is O or S;

[0021] Each R⁴ is independently selected from H, C1-C10 alkyl; C2-C10alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; aryl;R⁸, halo; haloalkyl; CF₃; SR⁵; OR⁵; OC(O)R⁵; NR⁵R⁵; NR⁵R⁶; COOR⁵; NO₂;CN; C(O)R⁵; C(O)C(O)R⁵; C(O)NR⁵R⁵; S(O)_(n)R^(5:) S(O)_(n)NR⁵R⁵;NR⁵C(O)NR⁵R⁵; NR⁵C(O)C(O)R⁵; NR⁵C(O)R⁵; NR⁵(COOR⁵); NR⁵C(O)R⁸;NR⁵S(O)_(n)NR⁵R⁵; NR⁵S(O)_(n)R⁵; NR⁵S(O)_(n)R⁸; NR⁵C(O)C(O)NR⁵R⁵;NR⁵C(O)C(O)NR⁵R⁶; C1-C10 alkyl substituted with 1-3 independent aryl, R⁷or R⁸; or C2-C10 alkenyl substituted with 1-3 independent aryl, R⁷ orR⁸;

[0022] Each R⁵ is independently H; C1-C10 alkyl; C2-C10 alkenyl; C2-C10alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; aryl; R⁹; haloalkyl;C1-C10 alkyl substituted with 1-3 independent aryl, R⁷ or R⁹ groups;C3-C10 cycloalkyl substituted with 1-3 independent aryl, R⁷ or R⁹groups; or C2-C10 alkenyl substituted with 1-3 independent aryl, R⁷ orR⁹;

[0023] Each R⁶ is independently C(O)R⁵, COOR⁵, C(O)NR⁵R⁵, or S(O)_(n)R⁵;

[0024] Each R⁷ is independently halo, CF₃, SR¹⁰, OR¹⁰, OC(O)R¹⁰,NR¹⁰OR¹⁰, NR¹⁰OR¹¹, NR¹¹R¹¹, COOR¹⁰, NO₂, CN, C(O)R¹⁰, OC(O)NR¹⁰R¹⁰,C(O)NR¹⁰R¹⁰ N(R¹⁰)C(O)R¹⁰, N(R¹⁰)(COOR¹⁰), S(O)_(n)NR¹⁰R¹⁰;

[0025] Each R⁸ is independently a 5-8 membered monocyclic, 8-12 memberedbicyclic, or 11-14 membered tricyclic ring system comprising 1-3heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9heteroatoms if tricyclic, said heteroatoms independently selected fromO, N, or S, which may be saturated or unsaturated, and wherein 0, 1, 2,3 or 4 atoms of each ring may be substituted by a substituentindependently selected from C1-C10 alkyl; C2-C10 alkenyl; C2-C10alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; aryl; R⁹; halo; sulfur;oxygen, CF₃; SR⁵; OR⁵; OC(O)R⁵; NR⁵R⁵; NR⁵R⁶; NR⁶R⁶; COOR⁵; NO₂; CN;C(O)R⁵; C(O)NR⁵R⁵; S(O)_(n)NR⁵R⁵; NR⁵C(O)NR⁵R⁵; NR⁵C(O)R⁹;NR⁵S(O)_(n)NR⁵R⁵; NR⁵S(O)_(n)R⁹; C1-C10 alkyl substituted with 1-3independent R⁷, R⁹ or aryl; or C2-C10 alkenyl substituted with 1-3independent R⁷, R⁹ or aryl;

[0026] Each R⁹ is independently a 5-8 membered monocyclic, 8-12 memberedbicyclic, or 11-14 membered tricyclic ring system comprising 1-3heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9heteroatoms if tricyclic, said heteroatoms independently selected fromO, N, or S, which may be saturated or unsaturated, and wherein 0, 1, 2or 3 atoms of each ring may be substituted by a substituentindependently selected from C1-C10 alkyl; C2-C10 alkenyl; C2-C10alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; halo; sulfur; oxygen;CF₃; SR¹⁰; OR¹⁰; NR¹⁰R¹⁰; NR¹⁰R¹¹; NR¹¹R¹¹; COOR¹⁰; NO₂; CN;S(O)_(n)R¹⁰; S(O)_(n)NR¹⁰R¹⁰; C(O)R¹⁰; or C(O)NR¹⁰R¹⁰;

[0027] Each R¹⁰ is independently H; C1-C10 alkyl; C2-C10 alkenyl; C2-C10alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; haloalkyl; C1-C10 alkyloptionally substituted with 1-3 independent C1-C10 alkyl, C2-C10alkenyl, C2-C10 alkynyl, C3-C10 cycloalkyl, C4-C10 cycloalkenyl, halo,CF₃, OR¹², SR¹², NR¹²R¹², COOR¹², NO₂, CN, C(O)R¹², C(O)NR¹²R¹²,NR¹²C(O)R¹², N(R¹²)(COOR¹²), S(O)_(n)NR¹²R¹², or OC(O)R¹²; or phenyloptionally substituted with 1-3 independent C1-C10 alkyl, C2-C10alkenyl, C2-C10 alkynyl, C3-C10 cycloalkyl, C4-C10 cycloalkenyl, halo,CF₃, OR¹², SR¹², NR¹²R¹², COOR¹², NO₂, CN, C(O)R¹², C(O)NR¹²R¹²,NR¹²C(O)R¹², N(R¹²)(COOR¹²), S(O)_(n)NR¹²R¹², or OC(O)R¹²;

[0028] Each R¹¹ is independently C(O)R¹⁰, COOR¹⁰, C(O)NR¹⁰R¹⁰ orS(O)_(n)R¹⁰;

[0029] Each R¹² is independently H; C1-C10 alkyl; C2-C10 alkenyl; C2-C10alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; C1-C10 alkylsubstituted with 1-3 independent C2-C10 alkenyl, C2-C10 alkynyl, C3-C10cycloalkyl, C4-C10 cycloalkenyl, halo, CF₃, OR¹³, SR¹³, NR¹³R¹³, COOR¹³,NO₂, CN, C(O)R¹³, C(O)NR¹³R¹³, NR¹³C(O)R¹³, or OC(O)R¹³; or phenyloptionally substituted with 1-3 independent C1-C10 alkyl, C2-C10alkenyl, C2-C10 alkynyl, C3-C10 cycloalkyl, C4-C10 cycloalkenyl, halo,CF₃, OR¹³, SR¹³, NR¹³R¹³, COOR¹³, NO₂, CN, C(O)R¹³, C(O)NR¹³R¹³,NR¹³C(O)R¹³, or OC(O)R¹³;

[0030] Each R¹³ is independently H; C1-C10 alkyl; C2-C10 alkenyl; C2-C10alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; C1-C10 alkyl optionallysubstituted with halo, CF₃, OR¹⁴, SR¹⁴, NR¹⁴R¹⁴, COOR¹⁴, NO₂, CN; orphenyl optionally substituted with halo, CF₃, OR¹⁴, SR¹⁴, NR¹⁴R¹⁴,COOR¹⁴, NO₂, CN;

[0031] Each R¹⁴ is independently H; C1-C10 alkyl; C3-C10 cycloalkyl orphenyl;

[0032] Each R¹⁵ is independently H; CF₃; CN; COOR⁵; or C1-C10 alkylsubstituted with 1-3 independent OR⁵, SR⁵, or NR⁵R⁵;

[0033] Each R¹⁶ is independently H, C1-C10 alkyl; C2-C10 alkenyl; C2-C10alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; aryl; R⁸; halo;haloalkyl; CF₃; COOR⁵; C(O)R⁵; C(O)C(O)R⁵; C(O)NR⁵R⁵; S(O)_(n)R^(5:)S(O)_(n)NR⁵R⁵; C1-C10 alkyl substituted with 1-3 independent aryl, R⁷ orR⁸; or C2-C10 alkenyl substituted with 1-3 independent aryl, R⁷ or R⁸;

[0034] Each R¹⁷ is independently H; C1-C10 alkyl; C2-C10 alkenyl; C2-C10alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; aryl; R⁸; halo;haloalkyl; CF₃; SR⁵; OR¹⁸; OC(O)R⁵; NR⁵R⁵; NR⁵R⁶; COOR⁵; NO₂; CN;C(O)R⁵; C(O)C(O)R⁵; C(O)NR⁵R⁵; S(O)_(n)R^(5:) S(O)_(n)NR⁵R⁵;NR⁵C(O)NR⁵R⁵; NR⁵C(O)C(O)R⁵; NR⁵C(O)R⁵; NR⁵(COOR⁵); NR⁵C(O)R⁸;NR⁵S(O)_(n)NR⁵R⁵; NR⁵S(O)_(n)R⁵; NR⁵S(O)_(n)R⁸; NR⁵C(O)C(O)NR₅R₅;NR⁵C(O)C(O)NR⁵R⁶; C1-C10 alkyl substituted with 1-3 independent aryl, R⁷or R⁸; or C1-C10 alkenyl substituted with 1-3 independent aryl, R⁷ orR⁸;

[0035] Each R¹⁸ is independently aryl; R⁸; C1-C10 alkyl substituted with1-3 independent aryl, CF₃, OC(O)R¹⁰, NHR¹⁹, NR¹⁰OR¹¹, NR¹¹R¹¹, COOR¹⁰,NO₂, CN, C(O)R¹⁰, OC(O)NR¹⁰R¹⁰, C(O)NR¹⁰R¹⁰, N(R¹⁰)C(O)R¹⁰, N(R¹⁰)(COOR¹⁰), S(O)_(n)NR¹⁰R¹⁰, or R⁸; or C2-C10 alkenyl substituted with 1-3independent aryl, CF₃, OC(O)R¹⁰, NHR¹⁹, NR¹⁰R¹¹, NR¹¹R¹¹, COOR¹⁰, NO₂,CN, C(O)R¹⁰, OC(O)NR¹⁰R¹⁰, C(O)NR¹⁰R¹⁰, N(R¹⁰)C(O)R¹⁰, N(R¹⁰) (COOR¹⁰),S(O)_(n)NR¹⁰R¹⁰, or R⁸;

[0036] Each R¹⁹ is independently C2-C10 alkenyl; C2-C10 alkynyl; C3-C10cycloalkyl; C4-C10 cycloalkenyl; aryl; R⁹; haloalkyl;

[0037] Each R²⁰ is independently NR⁵R¹⁶; OR⁵; SR⁵; or halo;

[0038] Each haloalkyl is independently a C1-C10 alkyl substituted withone or more halogen atoms, selected from F, Cl, Br, or I, wherein thenumber of halogen atoms may not exceed that number that results in aperhaloalkyl group;

[0039] Each aryl is independently a 6-carbon monocyclic, 10-carbonbicyclic or 14-carbon tricyclic aromatic ring system optionallysubstituted with 1-3 independent C1-C10 alkyl; C2-C10 alkenyl; C2-C10alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; R⁹; halo; haloalkyl;CF₃; OR¹⁰; SR¹⁰; NR¹⁰R¹⁰; NR¹⁰R¹¹; COOR¹⁰; NO₂; CN; C(O)R¹⁰;C(O)C(O)R¹⁰; C(O)NR¹⁰R¹⁰; N(R¹⁰)C(O)NR¹⁰R¹⁰; N(R¹⁰)C(O)R¹⁰;N(R¹⁰)S(O)_(n)R¹⁰; N(R¹⁰)(COOR¹⁰); NR¹⁰C(O)C(O)R¹⁰; NR¹⁰C(O)R⁹;NR¹⁰S(O)_(n)NR¹⁰R¹⁰; NR¹⁰S(O)_(n)R⁹; NR¹²C(O)C(O)NR¹²R¹²; S(O)_(n)R¹⁰;S(O)_(n)NR¹⁰R¹⁰; OC(O)R¹⁰; C1-C10 alkyl substituted with 1-3 independentR⁹, halo, CF₃, OR¹⁰, SR¹⁰, OC(O)R¹⁰, NR¹¹R¹¹, NR¹⁰R¹⁰, NR¹⁰R¹¹, COOR¹⁰,NO₂, CN, C(O)R¹⁰, OC(O)NR¹⁰R¹⁰, C(O)NR¹⁰R¹⁰, N(R¹⁰)C(O)R¹⁰, N(R¹⁰)(COOR¹⁰), S(O)_(n)NR¹⁰R¹⁰; R¹⁰; or C2-C10 alkenyl substituted with 1-3independent R⁹, halo, CF₃, OR¹⁰, SR¹⁰, OC(O)R¹⁰, NR¹¹R¹¹, NR¹⁰R¹⁰,NR¹⁰R¹¹, COOR¹⁰, NO₂, CN, C(O)R¹⁰, OC(O)NR¹⁰R¹⁰, C(O)NR¹⁰R¹⁰,N(R¹⁰)C(O)R¹⁰, N(R¹⁰) (COOR¹⁰), S(O)_(n)NR¹⁰R¹⁰;

[0040] Each heterocyclyl is independently a 5-8 membered nonaromaticmonocyclic, 8-12 membered nonaromatic bicyclic, or 11-14 memberednonaromatic tricyclic, ring system comprising 1-4 heteroatoms ifmonocyclic, 1-8 heteroatoms if bicyclic, or 1-10 heteroatoms iftricyclic, said heteroatoms independently selected from O, N, or S;

[0041] Each heteroaryl is independently a 5-8 membered aromaticmonocyclic, 8-12 membered aromatic bicyclic, or 11-14 membered aromatictricyclic ring system comprising 1-4 heteroatoms if monocyclic, 1-8heteroatoms if bicyclic, or 1-10 heteroatoms if tricyclic, saidheteroatoms independently selected from O, N, or S.

[0042] In alternate embodiments, the compounds are of the formula above,wherein each R¹ is independently NHR³, and each R² is independentlyNHR³; alternatively wherein each R¹ is independently NHR³, and each R²is independently one of the formulae:

[0043] alternatively wherein each R¹ is independently NHR³, wherein theR³ group in R¹ is heteroaryl substituted with 1-4 independent R⁴ on eachring, (and alternatively wherein at least one of said R⁴ is not H), andeach R² is independently one of the formulae:

[0044] wherein m is 0-3, alternatively m is 1 or 2, alternatively m is1;

[0045] alternatively wherein each R¹ is independently NHR³; wherein theR³ group in R¹ is pyrazolyl, triazolyl, imidazolyl, pyrrolyl, indolyl,or indazolyl, each substituted with 1-4 independent R⁴ on each ring,(and alternatively wherein at least one of said R⁴ is not H, andalternatively wherein at least one of said R⁴ is not H and no R⁴ may bemethyl), and each R² is independently one of the formulae:

[0046] wherein m is 0-3, alternatively m is 1 or 2, alternatively m is1;

[0047] alternatively wherein each R¹ is independently R³, and each R² isindependently NHR³;

[0048] alternatively wherein each R¹ is independently heterocyclylsubstituted with 1-4 independent R⁴ on each ring, (and alternativelywherein at least one of said R⁴ is not H), and each R² is independentlyNHR³, wherein each R¹ may not be1-alkyl-1,2,3,4-tetrahydroisoquinolin-2-yl (wherein alkyl is defined asmethyl, ethyl or propyl); alternatively wherein each R¹ is independentlyheterocyclyl substituted with 1-4 independent R⁴ on each ring, (andalternatively wherein at least one of said R⁴ is not H), and each R² isindependently one of the formulae:

[0049] wherein each R¹ may not be1-alkyl-1,2,3,4-tetrahydroisoquinolin-2-yl (wherein alkyl is defined asmethyl, ethyl or propyl);

[0050] alternatively wherein each R¹ is independently heterocyclylsubstituted with 1-4 independent R⁴ on each ring, (and alternativelywherein at least one of said R⁴ is not H), wherein said heterocyclylcomprises at least one nitrogen heteroatom and said heterocyclyl isattached at said nitrogen heteroatom;

[0051] alternatively wherein each R¹ is independently heterocyclylsubstituted with 1-4 independent R⁴on each ring, (and alternativelywherein at least one of said R⁴ is not H), wherein said heterocyclylcomprises at least one nitrogen heteroatom and said heterocyclyl isattached at said nitrogen heteroatom, and each R² is independently NHR³,wherein each R¹ may not be 1-alkyl-1,2,3,4-tetrahydroisoquinolin-2-yl(wherein alkyl is defined as methyl, ethyl or propyl);

[0052] alternatively wherein each R¹ is independently pyrrolylsubstituted with 1-4 independent R⁴ on each ring, (and alternativelywherein at least one of said R⁴ is not H), and each R² is independentlyNHR³;

[0053] alternatively wherein each R¹ is independently pyrazolylsubstituted with 1-4 independent R⁴ on each ring, (and alternativelywherein at least one of said R⁴ is not H), and each R² is independentlyNHR³;

[0054] alternatively wherein each R¹ is independently benzimidazolylsubstituted with 1-4 independent R⁴ on each ring, (and alternativelywherein at least one of said R⁴ is not H), and each R² is independentlyNHR³;

[0055] alternatively wherein each R¹ is independently heteroarylsubstituted with 1-4 independent R⁴ on each ring, (and alternativelywherein at least one of said R⁴ is not H), wherein said heteroarylcomprises at least one nitrogen heteroatom and said heteroaryl isattached at said nitrogen heteroatom, and said heteroaryl is notunsubstituted pyrrolyl;

[0056] alternatively wherein each R¹ is independently heteroarylsubstituted with 1-4 independent R⁴ on each ring, (and alternativelywherein at least one of said R⁴ is not H), wherein said heteroarylcomprises at least one nitrogen heteroatom and said heteroaryl isattached at said nitrogen heteroatom, and said heteroaryl is notunsubstituted pyrrolyl, and each R² is independently NHR³;

[0057] alternatively wherein each R¹ is independently heteroarylsubstituted with 1-4 independent R⁴ on each ring, (and alternativelywherein at least one of said R⁴ is not H), wherein said heteroarylcomprises at least one nitrogen heteroatom and said heteroaryl isattached at said nitrogen heteroatom, and said heteroaryl is notunsubstituted pyrrolyl, and each R² is independently one of theformulae:

[0058] alternatively wherein each R² is independently NHR³, and each R¹is independently of the formula:

[0059] alternatively wherein each R² is independently NHR³; and each R¹is independently of the formula:

[0060] alternatively wherein each R² is independently NHR³; and each R¹is independently of the formula:

[0061] alternatively wherein each R² is independently NHR³, and each R¹is independently of the formula:

[0062] alternatively wherein R² is independently NHR⁵;

[0063] alternatively wherein each R¹ is independently any one offollowing formulae:

[0064] and alternatively wherein R¹ is independently any of formulaeabove and R² is independently NHR⁵.

[0065] In an alternate embodiment, the compound is of any of theformulae herein, wherein R¹ is independently NHR³ and R² isindependently

[0066] wherein R⁴ is as defined herein and m is 0, 1, 2, or 3.

[0067] In an alternate embodiment, the compound is of any of theformulae herein, wherein R¹ is independently NHR³ and R² isindependently

[0068] wherein R⁴ is as defined herein and m is 0, 1, 2, or 3.

[0069] Alternate embodiments are those of any of the formulae hereinwherein each R³ is independently phenyl substituted with 1-4 independentR⁴, wherein at least one R⁴ is not H; and those of any of the formulaeherein wherein each R³ is independently heteroaryl substituted with 1-4independent R⁴, wherein at least one R⁴ is not H.

[0070] Alternate embodiments are those of any of the formulae hereinwherein each R¹ is independently phenyl substituted with 1-4 independentR⁴, wherein at least one R⁴ is not H; and those of any of the formulaeherein wherein each R¹ is independently heteroaryl substituted with 1-4independent R⁴, wherein at least one R⁴ is not H; and those of any ofthe formulae herein wherein each R¹ is independently heterocyclylsubstituted with 1-4 independent R⁴, wherein at least one R⁴ is not H.

[0071] Alternate embodiments are those of any of the formulae hereinwherein each R⁴ is independently C(O)NR⁵R⁵; or C1-C10 alkyl substitutedwith 1-3 independent C(O)NR⁵R⁵.

[0072] Alternate embodiments are those of any of the formulae hereinwherein each R⁴ is independently R⁸; alternatively wherein each R⁴ isindependently a 5-8 membered monocyclic saturated ring comprising 1-3heteroatoms, said heteroatoms independently selected from O, N, or S; oralternatively wherein each R⁴ is independently a 5-8 membered monocyclicsaturated ring comprising 1-3 heteroatoms, said heteroatomsindependently selected from O, N, or S, wherein 1, 2, or 3 atoms of eachring may be substituted by a substituent independently selected fromC1-C10 alkyl; C2-C10 alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10cycloalkenyl; aryl; R⁹; halo; sulfur; oxygen; CF₃; SR⁵; OR⁵; OC(O)R⁵;NR⁵R⁵; NR⁵R⁶; NR⁶R⁶; COOR⁵; NO₂; CN; C(O)R⁵; C(O)NR⁵R⁵; S(O)_(n)NR⁵R⁵;NR⁵C(O)NR⁵R⁵; NR⁵C(O)R⁹; NR⁵S(O)_(n)NR⁵R⁵; NR⁵S(O)_(n)R⁹; C1-C10 alkylsubstituted with 1-3 independent R⁷, R⁹ or aryl; or C2-C10 alkenylsubstituted with 1-3 independent R⁷, R⁹ or aryl.

[0073] Alternate embodiments are those of any of the formulae hereinwherein each R⁴ is independently C1-C10 alkyl substituted with 1-3independent R⁷; alternatively C1-C10 alkyl substituted with 1-3independent R⁸; or alternatively OR⁵ wherein each R⁵ is independentlyC1-C6 alkyl substituted with 1 independent R⁷ or R⁸.

[0074] Alternate embodiments are those of any of the formulae hereinwherein each R² is independently NHR³.

[0075] Alternate embodiments are those of any of the formulae hereinwherein each heteroaryl is independently a 5-6 membered monocylic ring;alternatively a 9-10 membered bicyclic ring; or alternatively a 13-14membered tricyclic ring.

[0076] Alternate embodiments are those of any of the formulae hereinwherein each heteroaryl is independently a 5-6 membered monocylic ringcomprising 1-3 heteroatoms, alternatively 1-2 heteroatoms, oralternatively 1 heteroatom; alternatively a 9-10 membered bicyclic ringcomprising 1-6 heteroatoms, alternatively 1-3 heteroatoms, alternatively1-2 heteroatoms, or alternatively 1 heteroatom; or alternatively a 13-14membered tricyclic ring comprising 1-6 heteroatoms, alternatively 1-3heteroatoms, alternatively 1-2 heteroatoms, or alternatively 1heteroatom.

[0077] Alternate embodiments are those of any of the formulae hereinwherein each heterocyclyl is independently a 5-6 membered monocylicring; alternatively a 9-10 membered bicyclic ring; or alternatively a13-14 membered tricyclic ring.

[0078] Alternate embodiments are those of any of the formulae hereinwherein each heterocyclyl is independently a 5-6 membered monocylic ringcomprising 1-3 heteroatoms, alternatively 1-2 heteroatoms, oralternatively 1 heteroatom; alternatively a 9-10 membered bicyclic ringcomprising 1-6 heteroatoms, alternatively 1-3 heteroatoms, alternatively1-2 heteroatoms, or alternatively 1 heteroatom; or alternatively a 13-14membered tricyclic ring comprising 1-6 heteroatoms, alternatively 1-3heteroatoms, alternatively 1-2 heteroatoms, or alternatively 1heteroatom.

[0079] Alternate embodiments are those of any of the formulae hereinwherein each R¹⁷ is independently C2-C10 alkenyl; C2-C10 alkynyl; C3-C10cycloalkyl; C4-C10 cycloalkenyl; aryl; R⁸; haloalkyl; CF₃; SR⁵; OR¹⁸;OC(O)R⁵; NR⁵R⁵; NR⁵R⁶; COOR⁵; NO₂; CN; C(O)R⁵; C(O)C(O)R⁵; C(O)NR⁵R⁵;S(O)_(n)R^(5:) S(O)_(n)NR⁵R⁵; NR⁵C(O)NR⁵R⁵; NR⁵C(O)C(O)R⁵; NR⁵C(O)R⁵;NR⁵(COOR⁵); NR⁵C(O)R⁸; NR⁵S(O)_(n)NR⁵R⁵R; NR⁵S(O)_(n)R⁵; NR⁵S(O)_(n)R⁸;NR⁵C(O)C(O)NR⁵R⁵; NR⁵C(O)C(O)NR⁵R⁶; C1-C10 alkyl substituted with 1-3independent aryl, R⁷ or R⁸; or C1-C10 alkenyl substituted with 1-3independent aryl, R⁷ or R⁸.

[0080] Alternate embodiments are those of any of the formulae hereinwherein each R¹⁸ is independently C1-C6 alkyl substituted with 1-3,alternatively 1-2, or alternatively 1 independent aryl, CF₃, OC(O)R¹⁰,NHR¹⁹, NR¹⁰OR¹¹, NR¹¹R¹¹, COOR¹⁰, NO₂, CN, C(O)R¹⁰, OC(O)NR¹⁰R¹⁰,C(O)NR¹⁰R¹⁰, N(R¹⁰)C(O)R¹⁰, N(R¹⁰) (COOR¹⁰), S(O)_(n)NR¹⁰R¹⁰ or R⁸

[0081] Alternate embodiments are those of any of the formulae hereinwherein each m is 0-3, alternatively m is 1 or 2, or alternatively m is1.

[0082] The invention also relates to methods of inhibiting enzyme orpolypeptide activity, particularly of an enzyme or polypeptide describedherein, such as a phosphoryl tranferase, or alternatively a kinase, in amammal comprising the step of administering to said mammal a compound ofany of the formulae described herein or a composition comprising acompound of any of the formulae described herein. In one embodiment, theinvention relates to a method of inhibiting phosphoryl transferase,alternatively kinase, activity in a mammal comprising the step ofadministering to said mammal a compound, or a composition comprising acompound, of any one of the formulae described herein. Preferably, themammal is a human.

[0083] In another embodiment, the invention relates to a method ofinhibiting enzyme activity in a mammal comprising the step ofadministering to said mammal a compound, or a composition comprising acompound, of any of the formulae described herein. Preferably, themammal is a human.

[0084] The invention also relates to methods of treating disease and/ordisease symptoms, particularly those mediated by an enzyme orpolypeptide described herein, such as phosphoryl transferase mediated,or kinase mediated, disease or disease symptoms, in a mammal comprisingthe step of administering to said mammal a compound of any of theformulae described herein or a composition comprising a compound of anyof the formulae described herein. Such diseases or disease symptoms aredescribed herein. “Kinase mediated” disease or disease symptoms refersto disease or disease symptoms in which kinase activity is involved. Inone embodiment, this invention relates to a method of treating diseaseor disease symptoms, particularly kinase mediated disease or diseasesymptoms, in a mammal comprising the step of administering to saidmammal a compound, or a composition comprising a compound, of any of theformulae described herein. Preferably, the mammal is a human.

[0085] In an alternate embodiment, this invention relates to a method oftreating disease or disease symptoms in a mammal comprising the step ofadministering to said mammal a compound, or a composition comprising acompound, of any of the formulae described herein. Preferably, themammal is a human.

[0086] In the compounds described herein, the term “halo” refers to anyradical of fluorine, chlorine, bromine or iodine. The terms “alkyl”,“alkenyl” and “alkynyl” refer to hydrocarbon chains that may bestraight-chain or branched-chain, containing the indicated number ofcarbon atoms. For example, C1-C10 indicates the group may have from 1 to10 (inclusive) carbon atoms in it. The terms “ring” and “ring system”refer to a ring comprising the delineated number of atoms, said atomsbeing carbon or, where indicated, a heteroatom such as nitrogen, oxygenor sulfur. The ring itself, as well as any substitutents thereon, may beattached at any atom that allows a stable compound to be formed. Theterm “nonaromatic” ring or ring system refers to the fact that at leastone, but not necessarily all, rings in a bicylic or tricyclic ringsystem is nonaromatic.

[0087] Leaving groups are species that may be detached from a moleculeduring a reaction and are known in the art. Examples of such groupsinclude, but are not limited to, halogen groups (e.g., I, Br, F, Cl),sulfonate groups (e.g., mesylate, tosylate), sulfide groups (e.g.,SCH₃), and the like. Nucleophiles are species that may be attached to amolecule during reaction and are known in the art. Examples of suchgroups include, but are not limited to, amines, Grignard reagents,anionic species (e.g., alkoxides, amides, carbanions) and the like.

[0088] In the methods described herein, said mammal is preferably ahuman. The inhibitors described herein, however, are useful ininhibiting kinase activity in human cells and useful in rodent (e.g.,murine) and other species used as surrogates for investigating activityin vitro and in vivo in humans and against human kinases. The inhibitorsdescribed herein are also usefutl for investigating inhibition andactivity of kinases originating from species other than humans.

[0089] The compounds and compositions described herein are useful forinhibition of kinase activity of one or more enzymes. Kinases include,for example, protein kinases (e.g., tyrosine, serone/threonine,histidine), lipid kinases (e.g., phosphatidylinositol kinases PI-3,PI-4) and carbohydrate kinases. Further information relating to kinasestructure, function and and their role in disease or disease symptoms isavailable at the Protein Kinase Resource web site(http://www.sdsc.edu/Kinases/pk_home.html). Kinases may be ofprokaryotic, eukaryotic, bacterial, viral, fungal or archaea origin.Specifically, the compounds described herein are useful as inhibitors oftyrosine, serine/threonine or histidine protein kinases, (includingcombinations or those of mixed specificity, that is for exmaple, thosethat phosphorylate both tyrosine and serine/threonine residues) or lipidkinases. Examples of kinases that are inhibited by the compounds andcompositions described herein and against which the methods describedherein are useful include, but are not limited to, LCK, IRK(=INSR=Insulin receptor), IGF-1 receptor, SYK, ZAP-70, IRAK1, BLK, BMX,BTK, FRK, FGR, FYN, HCK, ITK, LYN, TEC, TXK, YES, ABL, SRC, EGF-R(=ErbB-1), ErbB-2 (=NEU=HER2), ErbB-4, FAK, FGFIR (=FGR-1), FGF2R(=FGR-2), IKK-1 (=IKK-ALPHA=CHUK), IKK-2 (=IKK-BETA), MET (=c-MET), NIK,PDGF receptor ALPHA, PDGF receptor BETA, TIE1, TIE2 (=TEK), VEGFR1(=FLT-1), VEGFR2 (=KDR), FLT-3, FLT-4, KIT, CSK, JAK1, JAK2, JAK3, TYK2,RIP, RIP-2, LOK, TAK1, RET, ALK, MLK3, COT, TRKA, PYK2, Activin-likeKinases (Alk1-7), EPHA(1-8), EPHB(1-6), RON, GSK3(A and B), Ilk, PDK1,SGK, Fes, Fer, MatK, Ark(1-3), Plk(1-3), LimK(1 and 2), RhoK, Pak (1-3),Raf(A, B, and C), PknB, CDK(1-10), Chk(1 and 2), CamK(I-IV), CamKK, CK1,CK2, PKR, Jnk(1-3), EPHB4, UL13, ORF47, ATM, PKA (α, β, and γ), P38(α,β, and γ), Erk(1-3), PKB (including all PKB subtypes) (=AKT-1, AKT-2,AKT-3), and PKC (including all PKC subtypes). and all subtypes of thesekinases. The compounds and compositions of the invention are thereforealso particularly suited for treatment of diseases and disease symptomsthat involve one or more of the aforementioned protein kinases. In oneembodiment, the compounds, compositions or methods of this invention areparticularly suited for inhibition of or treatment of disease or diseasesymptoms mediated by LCK, ZAP, LYN, EGFR, ERB-B2, KDR, c-MET or SYK. Inanother embodiment, the compounds, compositions or methods of thisinvention are particularly suited for inhibition of or treatment ofdisease or disease symptoms mediated by src-family kinases. In anotherembodiment, the compounds, compositions or methods of this invention areparticularly suited for inhibition of or treatment of disease or diseasesymptoms mediated by kinases involved in angiogenesis. In anotherembodiment, the compounds, compositions or methods of this invention areparticularly suited for inhibition of or treatment of disease or diseasesymptoms mediated by kinases in one of the kinase families defined byHardie & Hanks, ed. supra., as in the the Src family (PTK-I), Syk/Zapfamily (PTK-VI), EGFR family (PTK-X), HGF Family (PTK-XXI), Insulinreceptor family (PTK-XVI), Tie/Tek family (PTK-XIII), Platelet-derivedgrowth factor receptor family (PTK-XIV), or Fibroblast growth factorreceptor family (PTK-XV). The compounds and compositions are also suitedfor regulating or modulating signal transduction in signal transductionpathways that involve one or more kinases, thus affecting events in acell, and are therefor useful in methods for regulating or modulatingsignal transduction.

[0090] The inhibitors described herein are also useful for inhibitingthe biological activity of any enzyme comprising greater than 90%,alternatively greater than 85%, or alternatively greater than 70%sequence homology with a phosphoryl transferase sequence, oralternatively a kinase sequence, including the kinases mentioned herein.The inhibitors described herein are also useful for inhibiting thebiological activity of any enzyme comprising a subsequence, or variantthereof, of any enzyme that comprises greater than 90%, alternativelygreater than 85%, or alternatively greater than 70% sequence homologywith a phosphoryl transferase subsequence, or alternatively kinasesubsequence, including subsequences of the kinases mentioned herein.Such subsequence preferably comprises greater than 90%, alternativelygreater than 85%, or alternatively greater than 70% sequence homologywith the sequence of an active site or subdomain of a phosphoryltransferase, or alternatively kinase, enzyme. The subsequences, orvariants thereof, comprise at least about 300, or alternatively at leastabout 200, amino acids.

[0091] The inhibitors described herein are useful for inhibiting thebiological activity of any enzyme that binds ATP and/or GTP and thus fortreating disease or disease symptoms mediated by any enzyme that bindsATP and/or GTP. The inhibitors described herein are also useful forinhibiting the biological activity of any enzyme that binds adenine orguanine nucleotides. The inhibitors described herein are also useful forinhibiting the biological activity of any enzyme that is involved inphosphotransfer and thus for treating disease or disease symptomsmediated by any enzyme that is involved in phosphotransfer. Theinhibitors described herein are also useful for inhibiting thebiological activity of a polypeptide or enzyme having sequence homologywith a phosphoryl transferase, or alternatively kinase, sequence andthus for treating disease or disease symptoms mediated by suchpolypeptide or enzyme. Such polypeptides or enzymes may be identified bycomparison of their sequence with phosphoryl transferase, alternativelykinase, sequences and phosphoryl transferase, alternatively kinase,catalytic domain sequences. Such sequences may be found, for example, indatabases such as GENEBANK, EMBO, or other similar databases known inthe art. For example, one method of comparison involves the databasePROSITE (http://expasy.hcuge.ch) (See, Hofmann K., Bucher P., FalquetL., Bairoch A., The PROSITE database, its status in 1999, Nucleic AcidsRes. 27:215-219(1999)), containing “signatures” or sequence patterns (ormotifs) or profiles of protein families or domains. Thus, the inhibitorsdescribed herein are useful for inhibiting the biological activity of apolypeptide or enzyme comprising a sequence that comprises a “signature”or sequence pattern or profile derived for, and identified in PROSITE asrelating to kinases, and for treating disease or disease symptomsmediated by such polypeptide or enzyme. Examples of such PROSITE motifsor consensus patterns identified as relating to kinases include PS00107,PS00108, PS00109, PS00112, PS00583, PS00584, PS50011, PS50290, PS00915,and PS00916.

[0092] The inhibitors described herein are also useful for inhibitingthe biological activity of ATP/GTP binding proteins. Many ATP/GTPbinding proteins have consensus motifs that can be used to identifythem. For example, PROSITE entry PDOC00017 titled “ATP/GTP-binding sitemotif A (P-loop)” describes a consensus pattern (called the A consensussequence or the P-loop) for a large group of nucleotide binding proteinsincluding ATP synthases, DNA and RNA helicases, ABC transporters,Kinesin and kinesin-like proteins, among many others. Other nucleotidebinding proteins have motifs similar to this P-loop, but take slightlydifferent forms. Examples of these include tubulins, lipid kinases andprotein kinases. The ATP binding motif of protein kinases have also beendefined within PROSITE entry PS00107. Yet other AGBPs have nothingsimilar to the P-loop motif. Examples of these include E1-E2 ATPases andthe glycolytic kinases.

[0093] The compounds, compositions and methods described herein areusefiul in inhibiting kinase activity. As such, the compounds,compositions and methods of this invention are useful in treatingkinase-mediated disease or disease symptoms in a mammal, particularly ahuman. Kinase mediated diseases are those wherein a protein kinase isinvolved in signaling, mediation, modulation, or regulation of thedisease process or symptoms. Kinase mediated diseases are exemplified bythe following disease classes: cancer, autoimmunological, metabolic,inflammatory, infection (bacterial, viral, yeast, fungal, etc.),diseases of the central nervous system, degenerative neural disease,allergy/asthma, dermatology, angiogenesis, neovascularization,vasculogenesis, cardiovascular, and the like.

[0094] The compounds, compositions and methods described herein areuseful in treating or preventing diseases or their symptoms, including,transplant rejection (e.g., kidney, liver, heart, lung, pancreas (isletcells), bone marrow, cornea, small bowel, skin allografts orxenografts), graft versus host disease, osteoarthritis, rheumatoidarthritis, multiple sclerosis, diabetes, diabetic retinopathy, asthma,inflammatory bowel disease (Crohn's disease, ulcerative colitis), renaldisease, cachexia, septic shock, lupus, diabetes mellitus, myastheniagravis, psoriasis, dermatitis, eczema, seborrhea, Alzheimer's disease,Parkinson's disease, stem cell protection during chemotherapy, ex vivoselection or ex vivo purging for autologous or allogeneic bone marrowtransplantation, leukemia (acute myeloid, chronic myeloid, acutelymphoblastic, etc.), cancer (breast, lung, colorectal, ovary, prostate,renal, squamous cell, prostate, glioblastoma, melanoma, pancreatic,Kaposi's sarcoma, etc.), ocular disease, retinopathies, (e.g., maculardegeneration, diabetic retinopathy), corneal disease, glaucoma,bacterial infections, viral infections, fungal infections and heartdisease, including but not limited to, restenosis. In one embodiment,the compositions and methods described herein are useful in treating orpreventing cancer, ocular disease, or retinopathies. In anotherembodiment, the compositions and methods described herein are useful intreating or preventing rheumatoid arthritis, transplant rejection,asthma or allergy, or their symptoms. In other embodiments, thecompositions and methods described herein are useful in treating orpreventing disease or disease symptoms involving hyperproliferativedisorders, or alternatively, involving angiogenesis.

[0095] Another embodiment envisioned by this invention relates to theuse of the kinase inhibitory compounds described herein for use asreagents that effectively bind to kinases. As reagents, the compounds ofthis invention, and their derivatives, may be derivatized to bind to astable resin as a tethered substrate for affinity chromatographyapplications. Such derivatives may be used in purification of enzymes,including phosphoryl transferases and kinases. The compounds of thisinvention, and their derivatives, may also be modified (e.g.,radiolabelled or affinity labelled, etc.) in order to utilize them inthe investigation of enzyme or polypeptide characterization, structure,and/or function. Additionally, the compounds described herein are usefulas reagents for chemical validation of drug targets. These and otheruses that characterize kinase inhibitors will be evident to those ofordinary skill in the art.

[0096] In another embodiment, the inhibitors described herein are usefulfor crystallizing or co-crystallizing with a protein kinase. Suchcrystals or crystal complexes may additionally comprise additionalpeptides and or metal ions. The crystals or crystal complexes may beused for investigation and determination of enzyme characteristicsincluding, for example, structure of the kinase enzyme, enzyme activesite domains, and inhibitor-enzyme interactions. This information isuseful in developing inhibitor compounds with modified characteristicsand for understanding structure-function relationships of the enzymesand their enzyme-inhibitor interactions.

[0097] In an alternate embodiment, the inhibitory compounds describedherein may be used as platforms or scaffolds which may be utilized incombinatorial chemistry techniques for preparation of derivatives and/orchemical libraries of compounds. Such derivatives and libraries ofcompounds have kinase inhibitory activity and are useful for identifyingand designing compounds possessing kinase inhibitory activity.Combinatorial techniques suitable for utilizing the compounds describedherein are known in the art as exemplified by Obrecht, D. andVillalgrodo, J. M., Solid-Supported Combinatorial and Parallel Synthesisof Small-Molecular-Weight Compound Libraries, Pergamon-Elsevier ScienceLimited (1998), and include those such as the “split and pool” or“parallel” synthesis techniques, solid-phase and solution-phasetechniques, and encoding techniques (see, for example, Czarnik, A. W.,Curr. Opin. Chem. Bio., (1997) 1, 60. Thus, one embodiment relates to amethod of using the compounds described in the formulae herein forgenerating derivatives or chemical libraries comprising: 1) providing abody comprising a plurality of wells; 2) providing one or more compoundsof the formulae described herein in each well; 3) providing anadditional one or more chemicals in each well; 4) isolating theresulting one or more products from each well. An alternate embodimentrelates to a method of using the compounds described in the formulaeherein for generating derivatives or chemical libraries comprising: 1)providing one or more compounds of the formulae described hereinattached to a solid support; 2) treating the one or more compounds ofthe formulae described herein attached to a solid support with one ormore additional chemicals; 3) isolating the resulting one or moreproducts from the solid support. In the methods described above, “tags”or identifier or labeling moieties may be attached to and/or detachedfrom the compounds of the formulae herein or their derivatives, tofacilitate tracking, identification or isolation of the desired productsor their intermediates. Such moieties are known in the art. Thechemicals used in the aforementioned methods may include, for example,solvents, reagents, catalysts, protecting group and deprotecting groupreagents and the like. Examples of such chemicals are those that appearin the various synthetic and protecting group chemistry texts andtreatises referenced herein.

[0098] The compounds of the formulae herein may be used to study themechanism and role of enzymes in biological pathways and processesinvolving kinases. The compounds of the formulae herein may also be usedas probes to identify new kinase enzymes or polypeptides with sequencehomology to kinases. The inhibitor compounds may be tethered to asupport or modified (e.g., tagged, radiolabeled or other identifiabledetection method) such that the compound may be detected and isolated inthe presence of the kinase enzyme or polypeptide. Thus, anotherembodiment relates to a method of identifying and/or isolating a kinaseenzyme or polypeptide with sequence homology to a kinase enzyme sequenceor subsequence, comprising, contacting a tethered or modified compoundof any of the formulae herein with one or more polypeptides, isolating apolypeptide/inhibitor complex, and identifying or isolating the sequenceof the polypeptide in the polypeptide/inhibitor complex. Theidentification of the polypeptide sequence may be performed while in thepolypeptide/inhibitor complex or after the polypeptide is decomplexedfrom the tethered or modified compound of any of the formulae herein.

[0099] The compounds are also useful in inhibiting enzymes, includingkinases, that play a role in plant metabolism regulation, plant growthor growth inhibition. As such the compounds and compositions of theinvention are useful as plant growth regulators, and as herbicides. Suchcompositions comprise the compounds of the invention as well as anyagricultural or other acceptable carrier for dispersal of the activecompound, such carriers and their use are known in the art.

[0100] Table 1 lists representative individual compounds of theinvention and compounds employed in the compositions and methods of thisinvention. TABLE 1  1  2

 3  4

 5  6

 7  8

 9 10

11 12

13 14

15 16

17 18

19 20

21 22

23 24

25 26

27 28

29 30

31 32

33 34

35 36

37 38

39 40

41 42

43 44

45 46

47 48

49 50

51 52

53 54

55 56

57 58

59 60

[0101] Combinations of substituents and variables envisioned by thisinvention are only those that result in the formation of stablecompounds. The term “stable”, as used herein, refers to compounds whichpossess stability sufficient to allow manufacture and which maintainsthe integrity of the compound for a sufficient period of time to beuseful for the purposes detailed herein (e.g., therapeutic orprophylactic administration to a mammal or for use in affinitychromatography applications). Typically, such compounds are stable at atemperature of 40° C. or less, in the absence of excessive moisture forat least one week.

[0102] As used herein, the compounds of this invention, including thecompounds of formulae described herein, are defined to includepharmaceutically acceptable derivatives or prodrugs thereof. A“pharmaceutically acceptable derivative or prodrug” means anypharmaceutically acceptable salt, ester, salt of an ester, or otherderivative of a compound of this invention which, upon administration toa recipient, is capable of providing (directly or indirectly) a compoundof this invention. Particularly favored derivatives and prodrugs arethose that increase the bioavailability of the compounds of thisinvention when such compounds are administered to a mammal (e.g., byallowing an orally administered compound to be more readily absorbedinto the blood) or which enhance delivery of the parent compound to abiological compartment (e.g., the brain or lymphatic system) relative tothe parent species. Preferred prodrugs include derivatives where a groupwhich enhances aqueous solubility or active transport through the gutmembrane is appended to the structure of formulae described herein.

[0103] Pharmaceutically acceptable salts of the compounds of thisinvention include those derived from pharmaceutically acceptableinorganic and organic acids and bases. Examples of suitable acid saltsinclude acetate, adipate, alginate, aspartate, benzoate,benzenesulfonate, bisulfate, butyrate, citrate, camphorate,camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate,ethanesulfonate, formate, fumarate, glucoheptanoate, glycerophosphate,glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride,hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate,oxalate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate,picrate, pivalate, propionate, salicylate, succinate, sulfate, tartrate,thiocyanate, tosylate and undecanoate. Other acids, such as oxalic,while not in themselves pharmaceutically acceptable, may be employed inthe preparation of salts useful as intermediates in obtaining thecompounds of the invention and their pharmaceutically acceptable acidaddition salts. Salts derived from appropriate bases include alkalimetal (e.g., sodium), alkaline earth metal (e.g., magnesium), ammoniumand N-(alkyl)₄ ⁺ salts. This invention also envisions the quaternizationof any basic nitrogen-containing groups of the compounds disclosedherein. Water or oil-soluble or dispersible products may be obtained bysuch quaternization.

[0104] The compounds of this invention may be synthesized usingconventional techniques. Advantageously, these compounds areconveniently synthesized from readily available starting materials. Ingeneral, the compounds of the formulae described herein are convenientlyobtained via methods illustrated in General Synthetic Schemes I-VIII andthe Examples herein. These general schemes are also exemplified by thespecific methods described in the Examples section below. GeneralSynthetic Schemes I-VIII and the examples utilize general chemical groupdescriptors (e.g., X, R³, R⁵) that are meant to be representative of anygroup suitable for synthesis of the compounds delineated herein. Suchgroups are exemplified by and include, but are not limited to, thosedefined in the definitions of the groups designated R¹, R², R³, R⁴, R⁵,R⁸, R¹², R¹⁶, R¹⁷, and R²⁰, for example, in the formulae herein. Citedreferences are incorporated by reference in their entirety.

[0105] Thus, one embodiment relates to a method of making a compound ofthe formulae described herein, comprising synthesizing any one or moreintermediates illustrated in the synthetic schemes herein and thenconverting that intermediate(s) to a compound of the formulae describedherein. Another embodiment relates to a method of making a compound ofthe formulae described herein, comprising synthesizing any one or moreintermediates illustrated in the examples herein and then convertingthat intermediate(s) to a compound of the formulae described herein.Another embodiment relates to a method of making a compound of theformulae described herein, comprising synthesizing any one or moreintermediates illustrated in the synthetic schemes herein and thenconverting that intermediate(s) to a compound of the formulae describedherein utilizing one or more of the chemical reactions described in thesynthetic schemes or examples herein. Nucleophilic agents are known inthe art and are described in the chemical texts and treatises referredto herein. The chemicals used in the aforementioned methods may include,for example, solvents, reagents, catalysts, protecting group anddeprotecting group reagents and the like. The methods described abovemay also additionally comprise steps, either before or after the stepsdescribed specifically herein, to add or remove suitable protectinggroups in order to ultimately allow synthesis of the compound of theformulae described herein.

[0106] In General Synthetic Scheme I, commercially availabledichloropyrimidine is sequentially treated, in the presence of base,with nucleophilic forms of R¹ and then R² to provide compounds of theinvention. Appropriate nucleophiles (e.g., HNRR, HSR, HOR, or theiranion equivalents, carbon anions, etc.) are known in the art.

[0107] In a similar fashion, this concept is illustrated in GeneralSynthetic Scheme II, wherein a benzimidazolyl derivative isrepresentative of R¹ and an amine derivative is representative of R².

[0108] General Synthetic Scheme III illustrates various pathways for thesynthesis of various compounds of the invention wherein one of R¹ or R²is a nitrogen attached heterocyclyl or heteroaryl group (represented asan optionally substituted R⁸).

[0109] General Synthetic Scheme IV illustrates an alternate method forconverting a leaving group-substituted pyrimidinyl intermediate (e.g., achloropyrimidine) to a corresponding aminopyrimidine compound usingacidic conditions. This alternative may be appropriate in place of anystep in a process depicted herein using basic conditions, as determinedby one of ordinary skill.

[0110] General Synthetic Scheme V illustrates one method forinterconversion of a leaving group on a pyrimidine core to an alternateleaving group. Such compounds are useful intermediates for synthesis ofcompounds of the formulae herein.

[0111] General Synthetic Scheme VI illustrates methods for conversion ofleaving group substituted pyrimidines to acyl- and alkyl-substitutedpyrimidines described herein.

[0112] General Synthetic Scheme VII illustrates a general method forsynthesis of aryl-substituted pyrimidines described herein.

[0113] General Synthetic Scheme VIII illustrates alternate methodologyfor synthesis of aryl-substituted pyrimidines described herein.

[0114] Alternatively, a compound of any of the formulae delineatedherein may be synthesized according to any of the processes delineatedherein. In the processes delineated herein, the steps may be performedin an alternate order and may be preceded, or followed, by additionalprotection/deprotection steps as necesssary. The processes may furthercomprise use of appropriate reaction inert solvents, additionalreagents, such as bases (e.g., LDA, diisopropylethylamine, pyridine,K₂CO₃, and the like), catalysts, and salt forms of the above. Theintermediates may be isolated or carried on in situ, with or withoutpurification. Purification methods are known in the art and include, forexample, crystallization, chromatography (liquid and gas phase,simulated moving bed (“SMB”)), extraction, distillation, trituration,reverse phase HPLC and the like. Reactions conditions such astemperature, duration, pressure, and atmosphere (inert gas, ambient) areknown in the art and may be adjusted as appropriate for the reaction.

[0115] Thus, one embodiment relates to a method of making a compound ofthe formulae described herein, comprising the step of reacting a mono-or di-leaving group substituted-1,3-pyrimidine, for example, a2-,4-dihalosubstituted-1,3-pyrimidine, with nucleophilic agents (e.g.,an aniline or amine) in 1 or 2 steps to form the compound of theformulae described herein. Nucleophilic agents are known in the art andare described in the chemical texts and treatises referred to herein.Such agents may have carbon or a heteroatom (e.g, N, O, S) as thenucleophilic atom. The chemicals used in the aforementioned methods mayinclude, for example, solvents, reagents, catalysts, protecting groupand deprotecting group reagents and the like. The methods describedabove may also additionally comprise steps, either before or after steps1 and 2 described above, to add or remove suitable protecting groups inorder to ultimately allow synthesis of the compound of the formulaedescribed herein.

[0116] In one embodiment, the invention relates to a process for makinga compound of any of the formulae described herein, comprising reactinga pyrimidine of one or more of the formulae:

[0117] with an appropriate nucleophilic agent or agents, wherein thegroups in said formulae are as defined herein.

[0118] In one embodiment, the invention relates to a process for makinga compound of any of the formulae described herein, comprising reactinga pyrimidine of one or more of the formulae:

[0119] with an appropriate nucleophilic agent or agents, wherein L isdefined as a leaving group and the groups in said formulae are asdefined herein.

[0120] In one embodiment, the invention relates to a process for makinga compound of the formula:

[0121] wherein

[0122] Each R¹ and R² is independently R³; R⁸; NHR³; NHR⁵; NHR⁶; NR⁵R⁵;NR⁵R⁶; SR⁵; SR⁶; OR⁵; OR⁶; C(O)R³; heterocyclyl optionally substitutedwith 1-4 independent R⁴ on each ring; or C1-C10 alkyl substituted with1-4 independent R⁴;

[0123] Each R³ is independently aryl; phenyl optionally substituted with1-4 independent R⁴; or heteroaryl optionally substituted with 1-4independent R⁴ on each ring; and all other substituents are as definedherein, or alternatively a compound of any one of the formulae describedherein,; comprising the steps of:

[0124] a) reacting a compound of formula (II) wherein each L isindependently a leaving group as defined herein, with a nucleophile offormula H—R¹ (or salt thereof) to give a compound of formula (III); and

[0125] b) reacting the compound of formula (III) with a nucleophile offormula H—R² (or salt thereof) to give a compound of formula (I).

[0126] In another embodiment, the process above is carried out byutilizing a nucleophile H—R² in step (a), then utilizing a nucleophileH—R¹ in step (b), as shown:

[0127] L is defined as a leaving group, and R¹ and R² are as definedherein.

[0128] In an alternate embodiment, the above-delineated processes areused to synthesize a compound of any of the formulae delineated herein.

[0129] As can be appreciated by the skilled artisan, the above syntheticschemes are not intended to comprise a comprehensive list of all meansby which the compounds described and claimed in this application may besynthesized. Further methods will be evident to those of ordinary skillin the art. Additionally, the various synthetic steps described abovemay be performed in an alternate sequence or order to give the desiredcompounds. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing theinhibitor compounds described herein are known in the art and include,for example, those such as described in R. Larock, Comprehensive OrganicTransformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 2d. Ed., John Wiley and Sons(1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995).

[0130] The compounds of this invention may be modified by appendingappropriate functionalities to enhance selective biological properties.Such modifications are known in the art and include those which increasebiological penetration into a given biological compartment (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion.

[0131] The novel compounds of the present invention are excellentligands for protein kinases, subsequences thereof, and homologouspolypeptides. Accordingly, these compounds are capable of targeting andinhibiting kinase enzyme and subsequences thereof. Inhibition can bemeasured by various methods, including, for example, those methodsillustrated in the examples below. The compounds described herein may beused in assays, including radiolabelled, antibody detection,calorimetric, and fluorometric, for the isolation, identification, orstructural or functional characterization of enzymes, peptides orpolypeptides. Other suitable assays include direct ATP competitiondisplacement assays where no phosphoryl transfer is necessary. Suchassays include any assay wherein a nucleoside or nucleotide arecofactors or substrates of the polypeptide of interest, and particularlyany assay involving phosphotransfer in which the substrates and orcofactors are ATP, GTP, Mg, Mn, peptides, polypeptides, lipids, orpolymeric amino acids.

[0132] Pharmaceutical compositions of this invention comprise a compoundof the formulae described herein or a pharmaceutically acceptable saltthereof; an additional agent selected from a kinase inhibitory agent(small molecule, polypeptide, antibody, etc.), an immunosuppressant, ananticancer agent, an anti-viral agent, antiinflammatory agent,antifungal agent, antibiotic, or an anti-vascular hyperproliferationcompound; and any pharmaceutically acceptable carrier, adjuvant orvehicle. Alternate compositions of this invention comprise a compound ofthe formulae described herein or a pharmaceutically acceptable saltthereof; and a pharmaceutically acceptable carrier, adjuvant or vehicle.Such compositions may optionally comprise one or more additionaltherapeutic agents, including, for example, kinase inhibitory agents(small molecule, polypeptide, antibody, etc.), immunosuppressants,anti-cancer agents, anti-viral agents, antiinflammatory agents,antifungal agents, antibiotics, or anti-vascular hyperproliferationcompounds.

[0133] The term “pharmaceutically acceptable carrier or adjuvant” refersto a carrier or adjuvant that may be administered to a patient, togetherwith a compound of this invention, and which does not destroy thepharmacological activity thereof and is nontoxic when administered indoses sufficient to deliver a therapeutic amount of the compound.

[0134] Pharmaceutically acceptable carriers, adjuvants and vehicles thatmay be used in the pharmaceutical compositions of this inventioninclude, but are not limited to, ion exchangers, alumina, aluminumstearate, lecithin, self-emulsifying drug delivery systems (SEDDS) suchas d-α-tocopherol polyethyleneglycol 1000 succinate, surfactants used inpharmaceutical dosage forms such as Tweens or other similar polymericdelivery matrices, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, potassium sorbate,partial glyceride mixtures of saturated vegetable fatty acids, water,salts or electrolytes, such as protamine sulfate, disodium hydrogenphosphate, potassium hydrogen phosphate, sodium chloride, zinc salts,colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,cellulose-based substances, polyethylene glycol, sodiumcarboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat. Cyclodextrins such as α-, β-, and γ-cyclodextrin, orchemically modified derivatives such as hydroxyalkylcyclodextrins,including 2- and 3-hydroxypropyl-β-cyclodextrins, or other solubilizedderivatives may also be advantageously used to enhance delivery ofcompounds of the formulae described herein.

[0135] The pharmaceutical compositions of this invention may beadministered orally, parenterally, by inhalation spray, topically,rectally, nasally, buccally, vaginally or via an implanted reservoir,preferably by oral administration or administration by injection. Thepharmaceutical compositions of this invention may contain anyconventional non-toxic pharmaceutically-acceptable carriers, adjuvantsor vehicles. In some cases, the pH of the formulation may be adjustedwith pharmaceutically acceptable acids, bases or buffers to enhance thestability of the formulated compound or its delivery form. The termparenteral as used herein includes subcutaneous, intracutaneous,intravenous, intramuscular, intraarticular, intraarterial,intrasynovial, intrasternal, intrathecal, intralesional and intracranialinjection or infusion techniques.

[0136] The pharmaceutical compositions may be in the form of a sterileinjectable preparation, for example, as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques known in the art using suitable dispersing or wetting agents(such as, for example, Tween 80) and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally acceptable diluent or solvent,for example, as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are mannitol, water, Ringer'ssolution and isotonic sodium chloride solution. In addition, sterile,fixed oils are conventionally employed as a solvent or suspendingmedium. For this purpose, any bland fixed oil may be employed includingsynthetic mono- or diglycerides. Fatty acids, such as oleic acid and itsglyceride derivatives are useful in the preparation of injectables, asare natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant, or carboxymethyl cellulose or similar dispersing agentswhich are commonly used in the formulation of pharmaceuticallyacceptable dosage forms such as emulsions and or suspensions. Othercommonly used surfactants such as Tweens or Spans and/or other similaremulsifying agents or bioavailability enhancers which are commonly usedin the manufacture of pharmaceutically acceptable solid, liquid, orother dosage forms may also be used for the purposes of formulation.

[0137] The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, emulsions and aqueous suspensions,dispersions and solutions. In the case of tablets for oral use, carrierswhich are commonly used include lactose and corn starch. Lubricatingagents, such as magnesium stearate, are also typically added. For oraladministration in a capsule form, useful diluents include lactose anddried corn starch. When aqueous suspensions and/or emulsions areadministered orally, the active ingredient may be suspended or dissolvedin an oily phase is combined with emulsifying and/or suspending agents.If desired, certain sweetening and/or flavoring and/or coloring agentsmay be added.

[0138] The pharmaceutical compositions of this invention may compriseformulations utilizing liposome or microencapsulation techniques. Suchtechniques are known in the art.

[0139] The pharmaceutical compositions of this invention may also beadministered in the form of suppositories for rectal administration.These compositions can be prepared by mixing a compound of thisinvention with a suitable non-irritating excipient which is solid atroom temperature but liquid at the rectal temperature and therefore willmelt in the rectum to release the active components. Such materialsinclude, but are not limited to, cocoa butter, beeswax and polyethyleneglycols.

[0140] Topical administration of the pharmaceutical compositions of thisinvention is especially useful when the desired treatment involves areasor organs readily accessible by topical application. For applicationtopically to the skin, the pharmaceutical composition should beformulated with a suitable ointment containing the active componentssuspended or dissolved in a carrier. Carriers for topical administrationof the compounds of this invention include, but are not limited to,mineral oil, liquid petroleum, white petroleum, propylene glycol,polyoxyethylene polyoxypropylene compound, emulsifying wax and water.Alternatively, the pharmaceutical composition can be formulated with asuitable lotion or cream containing the active compound suspended ordissolved in a carrier with suitable emulsifying agents. Suitablecarriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water. The pharmaceuticalcompositions of this invention may also be topically applied to thelower intestinal tract by rectal suppository formulation or in asuitable enema formulation. Topically-transdermal patches are alsoincluded in this invention.

[0141] The pharmaceutical compositions of this invention may beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other solubilizing or dispersingagents known in the art.

[0142] Dosage levels of between about 0.01 and about 100 mg/kg bodyweight per day, alternatively between about 0.5 and about 75 mg/kg bodyweight per day of the kinase inhibitory compounds described herein areuseful in a monotherapy and/or in combination therapy for the preventionand treatment of kinase mediated disease. Typically, the pharmaceuticalcompositions of this invention will be administered from about 1 toabout 6 times per day or alternatively, as a continuous infusion. Suchadministration can be used as a chronic or acute therapy. The amount ofactive ingredient that may be combined with the carrier materials toproduce a single dosage form will vary depending upon the host treatedand the particular mode of administration. A typical preparation willcontain from about 5% to about 95% active compound (w/w). Alternatively,such preparations contain from about 20% to about 80% active compound.

[0143] When the compositions of this invention comprise a combination ofa kinase inhibitor of the formulae described herein and one or moreadditional therapeutic or prophylactic agents, both the kinase inhibitorand the additional agent should be present at dosage levels of betweenabout 10 to 100%, and more preferably between about 10 to 80% of thedosage normally administered in a monotherapy regimen. The additionalagents may be administered separately, as part of a multiple doseregimen, from the compounds of this invention. Alternatively, thoseagents may be part of a single dosage form, mixed together with thecompounds of this invention in a single composition.

[0144] According to one embodiment, the pharmaceutical compositions ofthis invention may comprise an additional kinase inhibitory agent. Suchadditional kinase inhibitory agents are those which may modulate,regulate or otherwise affect kinase enzyme activity. Such effects maylead to modulation of disease pathology and/or symptoms. Kinaseinhibitory agents include, for example, small molecules, polypeptides,antibodies (including for example, monoclonals, chimeric, humanized,single chain, immunokines, etc.), and the like. Examples of additionalkinase inhibitory small molecule agents include, but are not limited to,SU-6668, SU-5416, ZD-4190, ZD-1839, STI-571, CP-358774, LY-333531 andthe like.

[0145] According to one embodiment, the pharmaceutical compositions ofthis invention comprise an additional immunosuppression agent. Examplesof additional immunosuppression agents include, but are not limited to,cyclosporin A, FK506, rapamycin, leflunomide, deoxyspergualin,prednisone, azathioprine, mycophenolate mofetil, OKT3, ATAG, interferonand mizoribine.

[0146] According to an alternate embodiment, the pharmaceuticalcompositions of this invention may additionally comprise antibodies(including for example, monoclonals, chimeric, humanized, single chain,immunokines, etc.), cytotoxic or hormonal anti-cancer agents orcombinations thereof. Examples of anti-cancer agents include, but arenot limited to, cis-platin, actinomycin D, doxorubicin, vincristine,vinblastine, etoposide, amsacrine, mitoxantrone, tenipaside, taxol,taxotere, colchicine, phenothiazines, interferons, thioxantheres,anti-estrogens (e.g., tamoxifen), aromatase inhibitors, anti-androgens,LHRH antagonists, progetins, and GnRH antagonists.

[0147] According to another alternate embodiment, the pharmaceuticalcompositions of this invention may additionally comprise an anti-viralagent. Examples of anti-viral agents include, but are not limited to,Cytovene, Ganciclovir, trisodium phosphonoformate, Ribavirin, d4T, ddl,AZT, amprenavir and acyclovir.

[0148] Upon improvement of a patient's condition, a maintenance dose ofa compound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level, treatment should cease.Patients may, however, require intermittent treatment on a long-termbasis upon any recurrence of disease symptoms.

[0149] As the skilled artisan will appreciate, lower or higher dosesthan those recited above may be required. Specific dosage and treatmentregimens for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health status, sex, diet, time ofadministration, rate of excretion, drug combination, the severity andcourse of the disease, condition or symptoms, the patient's dispositionto the disease, condition or symptoms, and the judgment of the treatingphysician.

[0150] In an alternate embodiment, this invention provides methods oftreating, preventing, or relieving symptoms of disease in a mammalcomprising the step of administrating to said mammal any of thepharmaceutical compositions and combinations described above.Preferably, the mammal is a human. If the pharmaceutical compositiononly comprises the inhibitor of this invention as the active component,such methods may additionally comprise the step of administering to saidmammal an additional therapeutic agent, such as an antiinflammatoryagent, immunosuppressant, an anti-cancer agent, an anti-viral agent, oran anti-vascular hyperproliferation compound. Such additional agent maybe administered to the mammal prior to, concurrently with, or followingthe administration of the inhibitor composition.

[0151] The compounds of this invention may contain one or moreasymmetric centers and thus occur as racemates and racemic mixtures,scalemic mixtures, single enantiomers, individual diastereomers anddiastereomeric mixtures. All such isomeric forms of these compounds areexpressly included in the present invention. The compounds of thisinvention may also be represented in multiple tautomeric forms, forexample, as illustrated below:

[0152] in such instances, the invention expressly includes alltautomeric forms of the compounds described herein. The compounds mayalso occur in cis- or trans- or E- or Z-double bond isomeric forms. Allsuch isomeric forms of such compounds are expressly included in thepresent invention. All crystal forms of the compounds described hereinare expressly included in the present invention.

[0153] Substituents on ring moieties (e.g., phenyl, thienyl, etc.) maybe attached to specific atoms, whereby they are intended to be fixed tothat atom, or they may be drawn unattached to a specific atom (seebelow), whereby they are intended to be attached at any available atomthat is not already substituted by an atom other than H (hydrogen). Forexample, a structure drawn as:

[0154] is intended to encompass all of the following structures:

[0155] The compounds of this invention may contain heterocyclic ringsystems attached to another ring system (e.g., a pyrimidinyl core ring,an R⁸ substituent as defined herein, or a heteroaryl group). Suchheterocyclic ring systems may be attached through a carbon atom or aheteroatom in the ring system. In instances wherein a heterocyclic orheteroaryl ring system is stated to be attached at a heteroatom (e.g.,nitrogen atom), this refers to the heterocyclic or heteroaryl ringsystem being attached to the designated functional group at saidnitrogen heteroatom. To illustrate, for example, when an R¹ or R²substituent on a pyrimidinyl core is a heteroaryl defined as beingattached at a nitrogen atom, this definition includes, but is notlimited to, structures such as those exemplified below:

[0156] All references cited herein, whether in print, electronic,computer readable storage media or other form, are expresslyincorporated by reference in their entirety, including but not limitedto, abstracts, articles, journals, publications, texts, treatises,internet web sites, databases, patents, and patent publications.

[0157] In order that the invention described herein may be more readilyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner. NMRand MS spectra obtained for compounds described in the examples belowand those described herein were consistent with that of the compounds ofthe formulae herein.

[0158] Analytical Methods:

[0159] Unless otherwise indicated all HPLC analyses are run on a HP-1050system with an HP Zorbax SB-C18 (5 μ) reverse phase column (4.6×150 mm)run at 30 degrees C. with a flow rate of 1.00 ml/minute.

[0160] The mobile phase used solvent A (water/0.1% trifluoroacetic acid)and solvent B (acetonitrile/0.1% trifluoroacetic acid) with a 20-minutegradient from 10% to 90% acetonitrile. The gradient is followed by a2-minute return to 10% acetonitrile and a 3 minute flush.

[0161] The peaks of interest eluted on the LC profiles at the timesindicated.

[0162] LC-MS Method:

[0163] 1. Samples are run on a HP-1100 MSD system with a HP Zorbax SB-C8(5 μ) reverse phase column (4.6×50 mm) run at 30 degrees C. with a flowrate of 0.75 ml/minute.

[0164] 2. The mobile phase used solvent A (water/0.1% acetic acid) andsolvent B (acetonitrile/0.1% acetic acid) with a 10-minute gradient from10% to 90% acetonitrile. The gradient is followed by a 1-minute returnto 10% acetonitrile and a 2 minute flush.

[0165] Proton NMR Spectra:

[0166] Unless otherwise indicated, all ¹H NMR spectra are run on aVarian series Mercury 300 MHz instrument. All observed protons arereported as parts-per-million (ppm) downfield from Tetramethylsilane(TMS) or other internal reference in the appropriate solvent indicated.

EXAMPLE 1

[0167]

[0168] The indole (10 mmole) is dissolved into DMF (20 mL) undernitrogen at room temperature, in a round bottom flask fitted with amagnetic stir bar and rubber septum. This solution is cooled to 0° C.with an ice-water bath. NaH (10 mmole, as the 60% suspension in mineraloil) is then added. Once gas evolution ceases, 2,4-dichloropyrimidine(10 mmole) is added as the solid. The reaction is then left to stirovernight with gradual warming to room temperature. Mass spectralanalysis of the crude reaction mixture shows complete reaction. Thereaction is quenched with saturated NH₄Cl_((aq)). This mixture is thendiluted with water and extracted with EtOAc (100 mL). The EtOAc extractsare then washed with water and brine, combined, dried over anhydrousNa₂SO₄, filtered, and concentrated under reduced pressure. The recoveredwaxey solid is then purified by flash silica gel column chromatography(5% and 10% EtOAc:Hexane step gradient) giving approximately 35% yield.

EXAMPLE 2

[0169]

[0170] The pyrimidine-indole substrate (0.5 mmol) is suspended intoisopropanol (6 mL) under air at rooom temperature in an open tube.Diisopropylethylamine (0.5 mmol) is added followed by addition of the3,4,5-trimethoxyaniline (0.5 mmol). The tube is then sealed and heatedto 100° C. overnight. The temperature of the reaction is graduallyincreased to 130° C. over 48 hours. The reaction is quenched by coolingit to room temperature. The solvent is removed under reduced pressureand the recovered solid is partially purified by flash silica gelchromatography (20%, 40%, 60%, 80% EtOAc:Hexane step gradient) givingrecovered unreacted pyrimidine-indole substrate (60%) and impure desiredproduct. The product is further purified by applying it to 500μprepplates and developing one time with a 7:7:7:1 MtBE:CH₂Cl₂:Hexane:MeOHeluant, followed by a methanol trituration of the recovered solid,giving an approximate 25% yield of an off-white solid.

EXAMPLE 3

[0171] Preparation of 3:

[0172] A solution of 0.36 g (2.0 mmol) of 3-t-butylacetate (prepared asin JOC, 1995, 60, 1565-1582) in 10 mL of DMF is cooled to 0° C., and tothis is added 0.075 g (2.2 mmol) of NaH (60% dispersion oil). Thereaction is stirred for 30 minutes, and then 0.3 g (2.0 mmol) of2,4-dichloropyrimidine is added as a solid. The ice bath is removed andthe reaction is stirred overnight at room temperature. The reaction isquenched with water, and the aqueous is extracted with 3×25 mL EtOAc.The combined organic layers are washed with brine and dried over MgSO₄.The crude product is purified by silica chromatography (hexane/ethylacetate, 4:1) to afford 0.06 g of 70: MS m/z=369 (M+Na).

[0173] To a solution of 0.032 g (0.2 mmol) of 3,4,5-trimethoxyaniline in5 mL of acetone is added 0.06 g (0.2 mmol) of 70, 5 drops of conc. HCland 0.5 mL of water. The reaction is heated to reflux and stirred for 12h. The reaction is then cooled. The resulting white precipitate isfiltered, washed with Et₂O and water and dried to afford 0.19 g of 3: MSm/z=394 (M+H); HPLC ret time=11.62 minutes.

EXAMPLE 4

[0174] Preparation of 11:

[0175] To a solution of 1.738 g (11.665 mmol) of 2,4-dichloropyrimidinein 30 ml DMF at 0° C. is added 2.03 ml (11.665 mmol) ofdiisopropylethylamine and 1.553 g (11.665 mmol) of 2-aminobenzimidazole.The reaction mixture is stirred at 40° C. for 4 days. The reaction isthen cooled to room temperature and diluted into water and ethylacetate. The layers are separated, and the organic layer is then washedthree times with brine, dried over sodium sulfate, and concentratedunder reduced pressure to yield 1.837 g of 71.

[0176] Intermediate 71 (264 mg=1.077 mmol) is combined with 197 mg(1.077 mmol) of 3,4,5-trimethoxyaniline and 0.188 ml (1.077 mmol) ofdiisopropylethylamine in 2 mL isopropyl alcohol. The mixture is heatedat about 120° C. overnight. The crude mixture is concentrated down underreduced pressure and purified on 2×1.0 mm silica gel prep plates with 5%methanol/dichloromethane as eluent to yield 81.7 mg (19%) of 11; MSm/z=393 (M+H); HPLC ret time: 10.37 minutes; ¹H NMR (DMSO-d₆)δ d 9.8 (s,1H), 8.3 (m, 2H), 7.7 (s, 2H), 7.0 (m, 3H), 6.7 (m, 2H), 6.6 (m, 1H),3.7 (s, 6H), 3.5 (s, 3H).

EXAMPLE 5

[0177] Preparation of 25:

[0178] To a solution of 0.2 g (1.5 mmol) of 2-benzoxazolinone in 5 mL ofDMF is added 0.050 g of NaH (60% dispersion oil). The reaction isstirred at room temperature for 30 min, and then a solution of 0.22 g of2,4-dichloropyrimidine in 1 mL of DMF is added. The reaction is stirredovernight and then quenched with water. The aqueous is extracted with3×25 mL of EtOAc, and combined organic extracts are washed with brineand dried over MgSO₄. The crude product is purified by silica gelchromotagraphy (hexane/EtOAc 4:1) to afford 0.046 g of 72 as an orangesolid; MS m/z=248 (M+H).

[0179] To a solution of 0.030 g (0.16 mmol) of 3,4,5-trimethoxyanilinein 10 mL of acetone is added 0.04 g (0.16 mmol) of 72, 3 drops of conc.HCl and 0.5 mL of water. The reaction is heated to reflux and stirredfor 14 h. The reaction is then cooled and evaporated. The orange residueis triturated with EtOAc and MeOH, and the resulting white precipitateis filtered, washed with MeOH and dried to afford 0.026 g of 25; MSm/z=395 (M+H).

EXAMPLE 6

[0180] Preparation of 31:

[0181] Indole (1 .15 g, 9.9 mmol) is dissolved into DMF (20 mL) under N₂and cooled to 0° C. NaH (404 mg of a 60% dispersion in mineral oil, 10.1mmol) is added, which produces a vigorous gas evolution. Once the gasevolution subsides 2,4-dichloropyrimidine (1.5 g, 10.1 mmol) is addedand the reaction is allowed to gradually warm to room temperatureovernight. The reaction is then quenched with saturated NH₄Cl_((aq)),diluted with water, and extracted three times with ethyl acetate. Theethyl acetate extracts are then washed with brine, combined, dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure. Therecovered material is then purified by elution through a 17×2.5 cmcolumn of silica gel (5% and 10% ethyl acetate:hexane step gradient)giving 793 mg (34%) of 73 as a white solid: MS m/z 230 [M+H]⁺; ¹H NMR(300 MHz, DMSO-d₆) δ 8.72 (d, J=5.9 Hz, 1H), 8.62 (d, J=8.0 Hz, 1H),8.21 (d, J=3.7 Hz, 1H), 7.95 (d, J=5.9 Hz, 1H), 7.68 (dd, J=7.7, 1.0 Hz,1H), 7.39 (m, 1H), 7.28 (t, J=7.7 Hz, 1H), 6.93 (d, J=3.7 Hz, 1H).

[0182] 2-Chloro-4-(1-indolyl)pyrimidine, 73, (121 mg, 0.53 mmol) issuspended into isopropanol (6 mL), under air at room temperature in atube. N,N-Diisopropylethylamine (68 mg, 0.53 mmol) is added, followed bythe addition of 3,4,5-trimethoxyaniline (97 mg, 0.53 mmol). The tube isthen sealed, and the reaction heated to 120° C. for 3 days. The reactionis then cooled to room temperature and concentrated under reducedpressure. The recovered material is then purified by elution through a17×2.5 cm column of silica gel (20%, 40%, 60% and 80% EtOAc: Hexane stepgradient) giving an impure brown solid that is then applied to two 500ii preparative TLC plates and developed one time with 7:7:7:1 MtBE:CH₂Cl₂: hexane: MeOH. The recovered material is then triturated withmethanol giving 50 mg (25%) of 31: MS m/z 377=[M+H]⁺; ¹H NMR (300 MHz,DMSO-d₆) δ 9.60 (s, 1H), 8.76 (br d, J=8.1 Hz, 1H), 8.49 (d, J=5.7 Hz,1H), 8.14 (d, J=3.4 Hz, 1H), 7.64 (d, J=7.4 Hz, 1H), 7.24 (m, 2H), 7.17(s, 2H), 6.82 (d, J=3.7 Hz, 1H), 5.76 (d, J=1.0 Hz, 1H), 3.74 (s, 6H),3.65 (s, 3H); HPLC Rt=11.54 min.

EXAMPLE 7

[0183] Preparation of 32:

[0184] To a solution of 2.0 g (13.4 mmol) of 2,4-dichloropyrimidine in25 mL of DMF is added 2.4 g (13.4 mmol) of 3,4,5-trimethoxyaniline and2.6 mL (14.7 mmol) of diisopropylethylamine. The mixture is heated to50° C. and stirred overnight. The reaction is quenched with water, sat.NH₄Cl and EtOAc, and the resulting precipitate is filtered and dried toafford 2.5 g of 74; MS m/z=296 (M+H); HPLC ret time=8.5 minutes; ¹H NMR(DMSO-d6) δ 10.0 (s, 1H), 8.1 (d, 1H), 6.9 (s, 2H), 6.7 (d, 1H), 3.7 (s,6H), 3.5 (s, 3H).

[0185] Indole (88 mg, 0.75 mmol) is dissolved into a 1:1 mixture of DMF:THF (5 mL). NaH (60 mg of a 60% dispersion in mineral oil, 1.5 mmol) isadded, which produces a vigorous gas evolution. Once the gas evolutionsubsides, 2-chloro-4-(3′,4′,5′-trimethoxyanilino)pyrimidine 74 (150 mg,0.5 mmol) is added, the tube is capped, and heated to 100° C. for twoweeks. The reaction is then cooled to room temperature and quenched withsaturated NH₄Cl_((aq)). This mixture is then diluted with water andextracted three times with ethyl acetate. The ethyl acetate extracts arethen washed with brine, combined, dried over anhydrous Na₂SO₄, filteredand concentrated under reduced pressure. The recovered material is thenpurified by elution through a 17×2.5 cm column of silica gel (20%, 40%,60% and 80% EtOAc: Hexane step gradient) giving an impure brown solidthat is then applied to two 500 μ preperative TLC plates and developedone time with 7:7:7:1 MtBE: CH₂Cl₂: hexane: MeOH giving 20 mg (10%) of32: MS m/z 377=[M+H]⁺; ¹H NMR (300 MHz, DMSO-d₆) δ 9.78 (s, 1H), 8.70(br d, J=7.7 Hz, 1H), 8.31 (d, J=6.0 Hz, 1H), 8.23 (d, J=3.4 Hz, 1H),7.62 (dd, J=5.9, 2.5 Hz, 1H), 7.19 (m, 2H), 6.97 (s, 2H), 6.74 (d, J=3.7Hz, 1H), 6.60 (d, J=6.0 Hz, 1H), 3.79 (s, 6H), 3.68 (s, 3H); HPLCRt=13.72 min.

EXAMPLE 8

[0186] Preparation of 33:

[0187] To a mixture of 0.20 g (1.1 mmol) of 3-(4-chlorophenyl)pyrazolein 5 mL of DMF is added 0.042 g of NaH (60% dispersion oil). Thereaction is stirred for 30 minutes at room temperature, and then asolution of 0.17 g (1.1 mmol) of 2,4-dichloropyrimidine in 1 mL of DMFis added. The reaction is stirred overnight, and then quenched withwater. The aqueous layer is extracted with 3×15 mL EtOAc, and thecombined organic layers are washed with brine and dried over MgSO₄. Theresulting precipitate is filtered and dried to afford 0.075 g of 75; ¹HNMR (DMSO-d6) δ 8.7 (d, 1H), 8.5 (d, 1H), 7.8 (m, 3H), 7.3 (m, 2H), 7.05(d, 1H).

[0188] To a solution of 0.037 g (0.25 mmol) of 3,4,5-trimethoxyanilinein 7 mL of acetone is added 0.073 g (0.25 mmol) of 75, 3 drops of conc.HCl and 2.0 mL of water. The reaction is heated to reflux and stirredfor 24 h. An additional 0.025 g of 3,4,5-trimethoxyaniline is added,along with 2 drops of conc HCl. The reaction is transferred to a sealedtube and is heated to 80° C. for 5 days. The reaction is then cooled andthe resulting precipitate is filtered, washed with water and dried toafford 0.011 g of 33; MS m/z=438 (M+H); HPLC ret time=17.2 minutes; ¹HNMR (DMSO-d6) δ 9.6 (s, 1H), 8.4 (m, 2H), 7.8 (m, 2H), 7.4 (m, 2H), 7.2(t, 1H), 7.05 (m, 1H), 7.0 (d, 2H) 3.7 (s, 6H), 3.5 (s, 3H).

EXAMPLE 9

[0189] Preparation of 34:

[0190] To a mixture of 0.20 g (1.1 mmol) of 3-(4-methoxyphenyl)pyrazolein 5 mL of DMF is added 0.042 g of NaH (60% dispersion oil). Thereaction is stirred for 30 minutes at room temperature, and then asolution of 0.17 g (1.1 mmol) of 2,4-dichloropyrimidine in 1 mL of DMFis added. The reaction is stirred overnight, and then quenched withwater. The aqueous layer is extracted with 3×15 mL EtOAc, and thecombined organic layers are washed with brine and dried over MgSO₄. Theresulting precipitate is filtered and dried to afford 0.13 g of 76; ¹HNMR (DMSO-d6) d 8.6 (t, 1H), 8.5 (m, 1H), 7.7 (m, 3H), 7.0 (t, 1H), 6.9(m, 2H), 3.6 (s, 3H).

[0191] To a solution of 0.097 g (0.65 mmol) of 3,4,5-trimethoxyanilinein 15 mL of acetone is added 0.124 g (0.4 mmol) of 76, 5 drops of conc.HCl and 2.0 mL of water. The reaction is heated to 120° C. in a sealedtube for 18 h. The reaction is then cooled and the resulting precipitateis filtered, washed with water and dried to afford 0.031 g of 34; MSm/z=434 (M+H); HPLC ret time=14.9 minutes; ¹H NMR (DMSO-d6) δ 9.5 (s,1H), 8.4 (d, 2H), 7.7 (m, 2H), 7.1 (m, 1H), 7.0 (d, 2H), 6.9 (s, 1H),6.8 (m, 2H), 3.5 (bs, 12H).

EXAMPLE 10

[0192] Preparation of 35:

[0193] To a solution of 0.20 g (1.3 mmol) of 2,4-dichloropyrimidine in 5mL of iPrOH is added 0.19 g (1.3 mmol) of 3-amino-5-t-butylpyrazole and0.25 mL (1.5 mmol) of diisopropylethylamine. The reaction is heated toreflux and is stirred for 10 h. The reaction is cooled and evaporated.The crude residue is purified by silica gel chromatography (5%MeOH/CH₂Cl₂) to afford 0.25 g of 77.

[0194] To a solution of 0.25 g (1.0 mmol) of 77 in 15 mL of acetone isadded 0.182 g (1.0 mmol) of 3,4,5-trimethoxyaniline, 3 drops of conc.HCl and 2.0 mL of water. The reaction is heated to reflux for 18 h. Thereaction is then cooled and the acetone is evaporated in vacuo. Theaqueous residue is extracted with EtOAc, and the organic layer is washedwith brine and dried over MgSO₄. The crude product is purified by silicagel chromatography (5% MeOH/CH₂Cl₂) to afford 0.29 g of 35: MS m/z=399(M+H); HPLC ret time=9.3 minutes; ¹H NMR (DMSO-d6) δ 11.8 (s, 1H), 9.3(s, 1H), 8.7 (s, 1H), 7.8 (d, 1H), 6.9 (s, 2H), 6.3 (s, 1H), 6.1 (s,1H), 3.6 (s, 6H), 3.4 (s, 3H).

EXAMPLE 11

[0195] Preparation of 36:

[0196] To a solution of 10 g (61.3 mmol) of 5-nitroindazole in 100 mL ofDMF is added 12.7 g (91.9 mmol) of K₂CO₃ and 7.29 mL (61.3 mmol) ofPhCH₂Br. The resulting mixture is stirred at RT for 3.5 days, and thenpoured into 400 mL of water. The resulting slurry is filtered, rinsedonce with water and dried in vacuo giving a beige solid. A 2.5 g portionof this crude material is purified by chromatography (SiO₂, elution with1:2 EtOAc-hexanes) giving 906.4 mg of 78.

[0197] To 906.4 mg (3.58 mmol) of 78 in 20 mL of MeOH and 5 mL of EtOAcat RT is added a slurry of 150 mg of 10% Pd—C in 5 mL of MeOH. Theresulting slurry is then stirred under a balloon of H₂ for 1.2 h, andfiltered through Celite™, rinsing with MeOH and EtOAc. Concentration ofthe filtrate gives 790.3 mg (98.9%) of 79 as a pinkish solid: MS m/z=224[M+H]⁺.

[0198] To a solution of 0.056 g (0.37 mmol) of 2,4-dichloropyrimidine in10 mL of isopropanol is added 0.084 g (0.37 mmol) of 79 and 0.07 mL ofdiisopropylethylamine (0.41 mmol). The reaction is heated to reflux andis stirred overnight. The organics are then removed in vacuo, and thecrude product is purified by silica chromatography (5% MeOH/CH₂Cl₂) toafford 0.12 g of 80.

[0199] To a solution of 0.12 g (0.36 mmol) of 80 in 10 mL of acetone isadded 0.066 g (0.036 mmol) of 3,4,5-trimethoxyaniline, 3 drops of conc.HCl and 1 mL of H₂O. The mixture is brought to reflux and is stirredovernight. The reaction is cooled and the acetone is evaporated. Theresulting oil is partitioned between EtOAc and water. The organicextracts are washed with brine and sat. NaHCO₃ and dried over MgSO₄. Thecrude product is purified by silica chromatography (5% MeOH/CH₂Cl₂) toafford 0.086 g of 36: MS m/z=483 (M+H); HPLC ret time=11.03 minutes; ¹HNMR (DMSO-d6) δ 9.1 (s, 1H), 8.7 (s, 1H), 8.05 (s, 1H), 7.7 (m, 2H), 7.4(d, 1H), 7.2 (d, 1H), 7.0 (m, 4H), 6.9 (s, 1H), 6.7 (d, 1H), 5.95 (d,1H), 5.4 (s, 2H), 3.4 (bs, 9H).

EXAMPLE 12

[0200] Preparation of 37:

[0201] To a solution of 0.071 g (0.24 mmol) of 74 in 10 mL of acetone isadded 0.054 g (0.24 mmol) of 79, 3 drops of HCl and 2 mL of H₂O. Thereaction is heated to reflux and is stirred for 30 h. The reaction isthen evaporated, and the resulting oil is partitioned between EtOAc andsat. NaHCO₃. The organic extracts are then washed with water, brine anddried over MgSO₄. The crude product is purified by silica chromatography(5% MeOH/CH₂Cl₂) to afford 0.053 g of 37: MS m/z=483 (M+H); HPLC rettime=11.4 minutes; ¹H NMR (DMSO-d6) δ 9.05 (s, 1H), 8.9 (s, 1H), 8.03(s, 1H), 7.79 (m, 1H), 7.7 (d, 1H), 7.3 (d, 2H), 7.1 (m, 2H), 7.0 (d,3H), 6.7 (s, 2H), 5.95 (d, 1H), 5.4 (s, 2H), 3.4 (bs, 9H).

EXAMPLE 13

[0202] Preparation of 54:

[0203] Compound 54 was prepared essentially by the method described inWO 97/19065 using the appropriate aniline reagents.

[0204] HPLC ret time=12.48 minutes; ¹H NMR (DMSO-d₆) δ 9.31 (s, 1H), 9.0(s, 1H), 7.9 (m, 1H), 7.75 (s, 1H), 7.55 (d, 1H), 7.32 (s, 2H), 7.07 (m,1H), 7.0 (d, 2H), 6.6 (d, 1H), 6.0 (d, 1H), 2.07 (s, 3H).

EXAMPLE 14

[0205] Preparation of 56:

[0206] Compound 56 was prepared essentially by the method described inWO 97/19065 using the appropriate aniline reagents.

[0207] MS m/z=383 (M+H); ¹H NMR (DMSO-d₆) δ 9.05 (s, 1H), 8.8 (s, 1H),7.9 (d, 1H), 7.5 (s, 1H), 7.15 (m, 2H), 6.85 (d, 1H), 6.75 (d, 1H), 6.05(d, 1H), 3.7 (2, 3H), 3.67 (s, 3H), 3.62 (s, 3H), 3.58 (s, 3H).

EXAMPLE 15

[0208] Preparation of 57:

[0209] Compound 57 was prepared essentially by the method described inWO 97/19065 using the appropriate aniline reagents.

[0210] MS m/z 331 (M+H); ¹H NMR (DMSO-d₆) δ 10.9 (s, 1H), 10.53 (s, 1H),8.0 (d, 1H), 7.72 (s, 1H), 7.65 (s, 1H), 7.52 (d, 1H), 7.39 (m, 3H),7.19 (m, 2H), 6.48 (d, 1H).

EXAMPLE 16

[0211] Preparation of 58:

[0212] Compound 58 was prepared essentially by the method described inWO 97/19065 using the appropriate aniline reagents.

[0213] HPLC ret time=12.70 minutes; ¹H NMR (DMSO-d₆) δ 9.12 (s, 1H),8.83 (s, 1H), 7.9 (d, 1H), 7.50 (d, 1H), 7.37 (m, 5H), 7.85 (m, 3H),7.81 (s, 1H), 6.1 (d, 1H), 5.0 (s, 2H), 3.65 (s, 6H), 3.58 (s, 3H).

EXAMPLE 17

[0214] Preparation of 59:

[0215] Compound 59 was prepared essentially by the method described inWO 97/19065 using the appropriate aniline reagents.

[0216]¹H NMR (DMSO-d₆) δ 9.2 (s, 1H), 9.11 (s, 1H), 7.92 (d, 1H), 6.68(d, 2H), 7.3 (t, 2H), 7.04 (t, 1H), 6.9 (m, 6H), 6.14 (d, 1H), 3.65 (s,6H), 3.56 (s, 3H).

EXAMPLE 18

[0217] Preparation of 60:

[0218] Compound 60 was prepared essentially by the method described inWO 97/19065 using the appropriate aniline reagents.

[0219] HPLC ret time=12.63 minutes; ¹H NMR (DMSO-d₆) δ 9.14 (s, 2H),7.85 (m, 2H), 7.5 (d, 1H), 7.33 (d, 1H), 7.23 (s, 1H), 7.0 (m, 2H), 6.85(d, 1H), 6.63 (d, 1H), 6.09 (d, 1H), 2.1 (s, 3H).

EXAMPLE 19

[0220] The inhibitor compounds described herein are screened in thefollowing manner. Kinases suitable for use in the following protocol todetermine kinase activity of the compounds described herein include, butare not limited to: Lck, Lyn, Src, Fyn, Syk, Zap-70, Itk, Tec, Btk,EGFR, ErbB2, Kdr, Flt-1, Flt-3, Tek, c-Met, InsR, and AKT.

[0221] Kinases are expressed as either kinase domains or fall lengthconstructs fused to glutathione S-transferase (GST) or polyHistidinetagged fusion proteins in either E. coli or Baculovirus-High Fiveexpression systems. They are purified to near homogeneity by affinitychromatography essentially as previously described (Lehr et al., 1996;Gish et al., 1995). In some instances, kinases are co-expressed or mixedwith purified or partially purified regulatory polypeptides prior tomeasurement of activity.

[0222] Kinase activity and inhibition are measured essentially byestablished protocols (Braunwalder et al., 1996). Briefly, The transferof ³³PO₄ from ATP to the synthetic substrates poly(Glu, Tyr) 4:1 orpoly(Arg, Ser) 3:1 attached to the bioactive surface of microtiterplates serves as the basis to evaluate enzyme activity. After anincubation period, the amount of phosphate transferred is measured byfirst washing the plate with 0.5% phosphoric acid, adding liquidscintillant, and then counting in a liquid scintillation detector. TheIC₅₀ is determined by the concentration of compound that causes a 50%reduction in the amount of ³³P incorporated onto the substrate bound tothe plate.

[0223] Other similar methods whereby phosphate is transferred to peptideor polypeptide substrate containing tyrosine, serine, threonine, orhistidine, either alone, in combination, or in combination with otheramino acids, in solution or immobilized (i.e., solid phase) are alsouseful. For example, transfer of phosphate to a peptide or polypeptidecan also be detected using scintillation proximity (Wu et al., 2000),ELISA (Cleaveland et al., 1990), Fluorescence Polarization (Seethala andMenzel, 1998), and homogeneous time-resolved fluorescence (HTRF, Kolb etal., 1998). Alternatively, kinase activity can be measured usingantibody-based methods whereby an antibody or polypeptide is used as areagent to detect phosphorylated target polypeptide. The compounds ofthe invention described herein are potent and selective kinaseinhibitors as demonstrated by representative compounds described hereinthat inhibit kinases with IC50 values at between about 10 nM and about 5M or greater. Representative results are summarized in the tables below.

REFERENCES

[0224] Braunwalder A F, Yarwood D R, Hall T, Missbach M, Lipson K E,Sills MA. (1996). A solid-phase assay for the determination of proteintyrosine kinase activity of c-src using scintillating microtitrationplates. Anal. Biochem. 234(1):23-26.

[0225] Cleaveland J S, Kiener P A, Hammond D J, Schacter B Z. (1990). Amicrotiter-based assay for the detection of protein tyrosine kinaseactivity. Anal Biochem. 190(2):249-53.

[0226] Gish G, McGlone M L, Pawson T, Adams J A. (1995). Bacterialexpression, purification and preliminary kinetic description of thekinase domain of v-fps. Protein Eng. 8(6):609-614.

[0227] Kolb, A. J., Kaplita, P. V., Hayes, D. J., Park, Y. -W., Pernell,C., Major, J. S., Mathis, G. (1998). Tyrosine kinase assays adapted tohomogeneous time-resolved fluorescence. Drug Discov. Today. 3:333-342.

[0228] Lehr R V, Ma Y G, Kratz D, Brake P G, Wang S, Faltynek C R, WangX M, Stevis P E (1996). Production, purification and characterization ofnon-myristylated human T-cell protein tyrosine kinase in a baculovirusexpression system. Gene 169(2):27527-9.

[0229] Seethala R, Menzel R. (1998). A fluorescence polarizationcompetition immunoassay for tyrosine kinases. Anal Biochem.255(2):257-62.

[0230] Wu J J, Yarwood D R, Sills M A, Chaudhuri B, Muller L, Zurini M,Sills M A. (2000). Measurement of cdk4 kinase activity using an affinitypeptide-tagging technology. Comb Chem High Throughput Screen.3(1):27-36.

EXAMPLE 20

[0231] The cellular activities of the inhibitor compounds describedherein may be assessed in a number of assays known to those skilled inthe art, some of which are exemplified as described below. Typicalsources for cells include, but are not limited to, human bone marrow orperipheral blood lymphocytes, fibroblasts, tumors, immortalized celllines, in-vitro transformed cell lines, rodent spleen cells, or theirequivalents. Tumor cells and transformed cell lines that have beenreported as cytokine- and growth factor-dependent cells are availablefrom standard cell banks such as The American Type Culture Collection(Bethesda, Md.). Cells genetically manipulated to express a particularkinase or kinases are also suitable for use in assaying cellularactivity and can be made using standard molecular biology methods. Thesecells are grown in various standard tissue culture media available fromsuppliers such as GIBCO/BRL (Grand Island, N.Y.) supplemented with fetalbovine serum. Cellular activity may also be measured using bacterial,yeast, or virally infected mammalian cells. Standard inhibitors (orreference compounds) of cellular activities measured in cellular assays,include mycophenolic acid (SIGMA, St. Louis, Mo.), staurosporine(Calbiochem, San Diego, Calif.), wortmannin (Calbiochem), cyclosporine,FK-506, and steroids (e.g., corticosteroids).

[0232] The compound(s) are tested for activity in cellular assays of Tor B cell activation. For example, the receptor-induced production ofcytokines and/or cell proliferation is a useful measure. This assay isperformed similarly to techniques described in the literature (1,2), andinvolves antibody-, antigen-, mitogen-, or antigen presentingcell-mediated crosslinking of the T cell or B cell receptor with orwithout engagement of co-stimulatory receptors.

[0233] The compound(s) are tested for activity in cellular assays ofallergic mediator release. For example, the receptor-induceddegranulation in mast cells or basophils leading to histamine releaseand the production of cytokines is a useful measure. This assay isperformed similarly to techniques described in the literature (3), andinvolves signalling via specific cell surface receptors for I, E, orother immunoglobulin (e.g., IgG) following crosslinking ofantigen-specific IgE on cells or immune complex binding leading todegranulation and or cytokine production.

[0234] The compound(s) are tested for activity in cellular assays ofgrowth factor effects. For example, growth factor receptor-inducedsignaling in a cell leading to intracellular signaling events such askinase autophosphorylation, phosphorylation of relevant kinasesubstrates, phosphorylation of MAP kinases, induction of geneexpression, or protein expression. Also, for example, growthfactor-induced functional events in cells such as DNA synthesis,proliferation, migration, or apoptosis. These assays are performedsimilarly to techniques described in the literature (4-7), and involveaddition of growth factor to responsive cells followed by monitoring ofsignaling or functional events.

[0235] The compound(s) are tested for activity in cellular assays oflymphokine, chemokine, cytokine, growth factor, or hormone, activation.For example, cytokine-induced intracellular signaling events and/or DNAsynthesis and/or cell proliferation and/or cytokine or chemokineproduction are a useful measure. These assays are performed similarly totechniques described in the literature (8), and involves addition ofcytokine to responsive cells followed by monitoring intracellularsignaling events and/or cell proliferation and/or cytokine production.

REFERENCES

[0236] 1. Shuji, K., et al. Activation of p21-CDC42/Rac-activatedkinases by CD28 signaling: p21-activated kinase (PAK) and MEK kinase 1(MEKK1) may mediate the interplay between CD3 and CD28 signals. J.Immunol. 160: 4182-4189 (1998).

[0237] 2. Satterthwaite, A. B., et al., Independent and opposing rolesfor Btk and Lyn in B cell and myeloid signaling pathways. J. Exp. Med.188: 833-844 (1998).

[0238] 3. Stephan, V., et al. FcεR1-induced protein tyrosinephosphorylation of pp72 in rat basophilic leukemia cells (RBL-2H3). J.Biol. Chem. 267 (8): 5434-5441 (1992).

[0239] 4. Olayioye, M. A., et al. ErbB-1 and ErbB-2 acquire distinctsignaling properties dependent upon their dimerization partner.Molecular and Cellular Biology. 18(9): 5042-5051 (1998).

[0240] 5. Buchdunger, E., et al. Inhibition of the Abl protein-tyrosinekinase in vitro and in vivo by a 2-phenylaminopyrimidine derivative.Cancer Res. 56;101-104 (1996).

[0241] 6. Yoshida, A. et al., Differential endothelial migration andproliferation to basic fibroblast growth factor and vascular endothelialgrowth factor. Growth Factors. 13:57-64 (1996).

[0242] 7. Brunet, A., et al., Akt promotes cell survival byphosphorylating and inhibiting a forkhead transcription factor. Cell.96:857-868 (1999).

[0243] 8. Liu, K. D., et al. Janus kinases in interleukin-2-mediatedsignaling: JAK1 and JAK3 are differentially regulated by tyrosinephosphorylation. Current Biology. 7 (11): 817-826 (1997).

[0244] Representative compounds tested under the following exampleprotocols exhibit cellular activities consistent with their observedenzyme inhibition activities.

EXAMPLE 21

[0245] Vascular Endothelial Growth Factor (VEGF)-Induced KdrAuto-Phosphorylation.

[0246] Human umbilical vein endothelial cells (HUVEC) are plated out inflat-well plates in complete media and allowed to adhere overnight. Thecells are then starved in medium containing 0.1% fetal calf serum (FCS),pre-incubated with or without dilutions of compound, then activated for15 minutes with 50 ng/ml VEGF. The cells are lysed and Kdr isimmunoprecipitated using an anti-Kdr antibody. The immunoprecipitatedKdr protein is separated by sodium dodecyl sulfate-polyacrylamide gelelectrophoresis (SDS-PAGE) and the level of phosphotyrosine isdetermined by western blotting with an anti-phosphotyrosine-specificantibody. IC₅₀'s are determined by comparing the level ofphosphotyrosine found in the presence of compound compared to controls.

EXAMPLE 22

[0247] Vascular Endothelial Growth Factor (VEGF)-Induced Extra-CellularSignal Regulated Kinase (Erk) 1/2-Phosphorylation.

[0248] Human umbilical vein endothelial cells (HUVEC) are plated out inflat-well plates in complete media and allowed to adhere overnight. Thecells are then starved in medium containing 0.1% fetal calf serum (FCS),pre-incubated with or without dilutions of compound, then activated for15 minutes with 50 ng/ml VEGF. The cells are lysed and proteins areseparated by SDS-PAGE. The level of phosphotyrosine on Erk1/2 isdetermined by western blotting with an anti-phospho-Erk1/2-specificantibody. IC₅₀'s are determined by comparing the level ofphosphotyrosine found in the presence of compound compared to controls.

EXAMPLE 23

[0249] Vascular Endothelial Growth Factor (VEGF)-Induced Proliferation.

[0250] Human umbilical vein endothelial cells (HUVEC) are plated out inflat-well plates in complete media and allowed to adhere overnight. Thecells are then starved in medium containing 0.1% fetal calf serum (FCS),pre-incubated with or without dilutions of compound, then activated for72 hours with 50 ng/ml VEGF. Proliferation is determined by the level of³H-thymidine incorporation into DNA. IC₅₀'s are determined by comparingthe level of thymidine incorporation found in the presence of compoundcompared to controls.

EXAMPLE 24

[0251] Growth Factor-Induced DNA Synthesis.

[0252] A rat fibroblast cell line is plated out in flat-well plates incomplete medium and allowed to adhere overnight. The cells are thenstarved in medium containing 0.1% bovine serum albumin (BSA),pre-incubated with or without dilutions of compound, then activatedovernight with 50 ng/ml platelet derived growth factor (PDGF), 1 ng/mlepidermal growth factor (EGF), 3 ng/ml fibroblast growth factor (FGF),or 10 ng/ml insulin-like growth factor-1 (IGF-1). Proliferation isdetermined by the level of ³H-thymidine incorporation into DNA. IC₅₀'sare determined by comparing the level of thymidine incorporation foundin the presence of compound compared to controls.

EXAMPLE 25

[0253] Platelet-Derived Growth Factor (PDGF)-Induced PDGF Receptor(PDGF-R) Auto-Phosphorylation.

[0254] A mouse fibroblast cell line is plated out in flat-well plates incomplete medium and allowed to adhere overnight. The cells are thenstarved in mediun containing 0.1% bovine serum albumin (BSA),pre-incubated with or without dilutions of compound, then activated with50 ng/ml platelet derived growth factor (PDGF) for 5 minutes. The cellsare lysed and proteins are separated by SDS-PAGE. The level ofphosphotyrosine on PDGF-R is determined by western blotting with ananti-phosphotyrosine-specific antibody. IC₅₀'s are determined bycomparing the level of phosphotyrosine found in the presence of compoundcompared to controls.

EXAMPLE 26

[0255] Epidermal Growth Factor (EGF)-Induced EGF Receptor (EGF-R)Auto-Phosphorylation.

[0256] Human epidermoid carcinoma cells (A431) are plated out inflat-well plates in complete media and allowed to adhere overnight. Thecells are then starved in medium containing 0.5% fetal calf serum (FCS),pre-incubated with or without dilutions of compound, then activated for3 minutes with 50 ng/ml EGF. The cells are lysed and proteins areseparated by SDS-PAGE. The level of phosphotyrosine on EGF-R isdetermined by western blotting with an anti-phospho-EGF-R-specificantibody. IC₅₀'s are determined by comparing the level ofphosphotyrosine found in the presence of compound compared to controls.

EXAMPLE 27

[0257] Heregulin-β1 (HRG)-Induced ErbB2 Auto-Phosphorylation.

[0258] Human breast carcinoma cells (ZR-75) are plated out in flat-wellplates in complete media and allowed to adhere overnight. The cells arethen starved in medium containing 0.5% fetal calf serum (FCS),pre-incubated with or without dilutions of compound, then activated for5 minutes with 50 ng/ml HRG. The cells are lysed and proteins areseparated by SDS-PAGE. The level of phosphotyrosine on ErbB2 isdetermined by western blotting with an anti-phospho-ErbB2-specificantibody. IC₅₀'s are determined by comparing the level ofphosphotyrosine found in the presence of compound compared to controls.

EXAMPLE 28

[0259] Hepatocyte Growth Factor (HGF) Receptor Met)Auto-Phosphorylation.

[0260] Human gastric carcinoma cells (MKN-45), which overexpress andconstitutively auto-phosphorylate Met, are plated out in flat-wellplates in complete media and allowed to adhere overnight. The cells arethen incubated with or without dilutions of compound for 1 hour. Thecells are lysed and proteins are separated by SDS-PAGE. The level ofphosphotyrosine on Met is determined by western blotting with ananti-phospho-tyrosine-specific antibody. IC₅₀'s are determined bycomparing the level of phosphotyrosine found in the presence of compoundcompared to controls.

EXAMPLE 29

[0261] Anti-CD3/CD28-Induced IL-2 Secretion and Proliferation.

[0262] Purified T cells are obtained from human peripheral bloodlymphocytes. T cells are pre-incubated incubated with or withoutdilutions of compound for 30 minutes. The T cells and compounds are thentransferred to a plate containing captured anti-CD3-specific antibody.Anti-CD28-specific antibody is then added and the cells are incubatedfor 20 hours. T cell supernatants are measured for the presence ofinterleukin-2 by commercially available ELISA. IC₅₀'s are determined bycomparing the level of IL-2 secretion found in the presence of compoundcompared to controls. The cells are then pulsed with ³H-thymidine andincubated for an additional 24 hours to determine cellularproliferation. IC₅₀'s are determined by comparing the level of thymidineincorporation found in the presence of compound compared to controls.

EXAMPLE 30

[0263] Anti-CD3-Induced T Cell Receptor ζ-chain (TCRζ) Phosphorylation.

[0264] The human T cell line, Jurkat, is pre-incubated with or withoutcompounds, then incubated with anti-CD3-specific antibody at 4° C. Cellsare washed, then incubated at 4° C. with a secondary anti-immunoglobulinantibody for crosslinking. Cells are activated by transfer to a 37° C.water bath for 1 minute. The cells are lysed and proteins are separatedby SDS-PAGE. The level of phosphotyrosine on TCRζ is determined bywestern blotting with an anti-phospho-tyrosine-specific antibody. IC₅₀'sare determined by comparing the level of phosphotyrosine found in thepresence of compound compared to controls.

[0265] The following tables summarize results (IC₅₀) of representativecompounds of the formulae described herein in assay protocols describedin Example 19. TABLE 2 Compound number Akt3-1 EGFR-1 ErbB2-1 ErbB4-1FGFR1-1 3 D D D D A 11 C D ND ND ND 25 D D D ND ND 31 D D ND B A 32 D DND ND ND 33 D D D ND ND 34 D C D ND ND 35 D C D C C 36 D A A ND ND 37 DB A ND ND 54 D D D ND ND 56 D D D ND B 57 D D D ND C 58 ND D ND ND ND 59ND C ND ND ND 60 D D ND ND ND

[0266] TABLE 3 Compound number Flt1-1 Fyn-1 Hck-1 IGFR-1 InsR-1 3 A C DA B 11 ND ND ND ND ND 25 ND ND ND D ND 31 A A D ND ND 32 ND ND ND ND ND33 ND ND ND D ND 34 ND ND ND D ND 35 C B ND A A 36 ND ND ND A ND 37 NDND ND A ND 54 ND ND ND A ND 56 B C ND A D 57 C ND ND A C 58 ND ND ND NDND 59 ND ND ND ND ND 60 ND ND ND A ND

[0267] TABLE 4 Compound number Itk-1 KDR-1 Lck-1 Lck-2 Lyn-1 3 D A B B B11 C C C B ND 25 D D D D ND 31 B A A A A 32 ND C D D ND 33 D D D D ND 34D D D D ND 35 B B A A A 36 A A A A ND 37 B B A A ND 54 D C D D ND 56 D AC A ND 57 C A D A ND 58 C C C A ND 59 C C C A ND 60 B B D A ND

[0268] TABLE 5 Compound number Met-1 PDGFRB Ret-1 Src-1 Tek-1 Zap-1 3 BA A C C B 11 C ND ND ND D D 25 D D ND ND D D 31 B A ND A C D 32 D ND NDND D D 33 D D ND ND ND C 34 D D ND ND ND C 35 A B B A ND D 36 B A ND NDND C 37 B A ND ND ND D 54 D C ND ND ND ND 56 D A B B ND D 57 B C A ND NDD 58 C ND ND ND ND ND 59 C ND ND ND ND ND 60 C B ND ND ND ND

[0269] While we have described a number of embodiments of thisinvention, it is apparent that our basic examples may be altered toprovide other embodiments that utilize the products and processes ofthis invention. Therefore, it will be appreciated that the scope of thisinvention is to be defined by the claims rather than by the specificembodiments that have been represented by way of example.

What is claimed is:
 1. A compound of the formula:

wherein, Each R¹and R² is independently R³; R⁸; NHR³; NHR⁵; NHR⁶; NR⁵R⁵;NR⁵R⁶; SR⁵; SR⁶; OR⁵; OR⁶; C(O)R³; heterocyclyl optionally substitutedwith 1-4 independent R⁴ on each ring; or C1-C10 alkyl substituted with1-4 independent R⁴; Each R³ is independently aryl; phenyl optionallysubstituted with 1-4 independent R⁴; or heteroaryl optionallysubstituted with 1-4 independent R⁴ on each ring; Each m isindependently 0, 1, 2 or 3; Each n is independently 1 or 2; Each X is Oor S; Each R⁴ is independently selected from H, C1-C10 alkyl; C2-C10alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; aryl;R⁸, halo; haloalkyl; CF₃; SR⁵; OR⁵; OC(O)R⁵; NR⁵R⁵; NR⁵R⁶; COOR⁵; NO₂;CN; C(O)R⁵; C(O)C(O)R⁵; C(O)NR⁵R⁵; S(O)_(n)R⁵: S(O)_(n)NR⁵R⁵;NR⁵C(O)NR⁵R⁵; NR⁵C(O)C(O)R⁵; NR⁵C(O)R⁵; NR⁵(COOR⁵); NR⁵C(O)R⁸;NR⁵S(O)_(n)NR⁵R⁵; NR⁵S(O)_(n)R⁵; NR⁵S(O)_(n)R⁸; NR⁵C(O)C(O)NR⁵R⁵;NR⁵C(O)C(O)NR⁵R⁶; C1-C10 alkyl substituted with 1-3 independent aryl, R⁷or R⁸; or C2-C10 alkenyl substituted with 1-3 independent aryl, R⁷ orR⁸; Each R⁵ is independently H; C1-C10 alkyl; C2-C10 alkenyl; C2-C10alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; aryl; R⁹; haloalkyl;C1-C10 alkyl substituted with 1-3 independent aryl, R⁷ or R⁹ groups;C3-C10 cycloalkyl substituted with 1-3 independent aryl, R⁷ or R⁹groups; or C2-C10 alkenyl substituted with 1-3 independent aryl, R⁷ orR⁹; Each R⁶ is independently C(O)R⁵, COOR⁵, C(O)NR⁵R⁵, or S(O)_(n)R⁵;Each R is independently halo, CF₃, SR¹⁰, OR¹⁰, OC(O)R¹⁰, NR¹⁰OR¹⁰,NR¹⁰OR¹¹, NR¹¹R¹¹, COOR¹⁰, NO₂, CN, C(O)R¹⁰, OC(O)NR¹⁰R¹⁰, C(O)NR¹⁰R¹⁰ ,N(R¹⁰)C(O)R¹⁰, N(R¹⁰)(COOR¹⁰), S(O)_(n)NR¹⁰R¹⁰; Each R⁸ is independentlya 5-8 membered monocyclic, 8-12 membered bicyclic, or 11-14 memberedtricyclic ring system comprising 1-3 heteroatoms if monocyclic, 1-6heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, saidheteroatoms independently selected from O, N, or S, which may besaturated or unsaturated, and wherein 0, 1, 2, 3 or 4 atoms of each ringmay be substituted by a substituent independently selected from C1-C10alkyl; C2-C10 alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10cycloalkenyl; aryl; R⁹; halo; sulfur; oxygen; CF₃; SR⁵; OR⁵; OC(O)R⁵;NR⁵R⁵; NR⁵R⁶; NR⁶R⁶; COOR⁵; NO₂; CN; C(O)R⁵; C(O)NR⁵R⁵; S(O)_(n)NR⁵R⁵;NR⁵C(O)NR⁵R⁵; NR⁵C(O)R⁹; NR⁵S(O)_(n)NR⁵R⁵; NR⁵S(O)_(n)R⁹; C1-C10 alkylsubstituted with 1-3 independent R⁷, R⁹ or aryl; or C2-C10 alkenylsubstituted with 1-3 independent R⁷, R⁹ or aryl; Each R⁹ isindependently a 5-8 membered monocyclic, 8-12 membered bicyclic, or11-14 membered tricyclic ring system comprising 1-3 heteroatoms ifmonocyclic, 1-6 heteroatoms if bicyclic, or 1-9 heteroatoms iftricyclic, said heteroatoms independently selected from O, N, or S,which may be saturated or unsaturated, and wherein 0, 1, 2 or 3 atoms ofeach ring may be substituted by a substituent independently selectedfrom C1-C10 alkyl; C2-C10 alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl;C4-C10 cycloalkenyl; halo; sulfur; oxygen; CF₃; SR¹⁰; OR¹⁰; NR¹⁰R¹⁰;NR¹⁰R¹¹; NR¹¹R¹¹; COOR¹⁰; NO₂; CN; S(O)_(n)R¹⁰; S(O)_(n)NR¹⁰R¹⁰;C(O)R¹⁰; or C(O)NR¹⁰R¹⁰; Each R¹⁰ is independently H; C1-C10 alkyl;C2-C10 alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl;haloalkyl; C1-C10 alkyl optionally substituted with 1-3 independentC1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C3-C10 cycloalkyl, C4-C10cycloalkenyl, halo, CF₃, OR¹², SR¹², NR¹²R¹², COOR¹², NO₂, CN, C(O)R¹²,C(O)NR¹²R¹², NR¹²C(O)R¹², N(R¹²)(COOR¹²), S(O)_(n)NR¹²R¹², or OC(O)R¹²;or phenyl optionally substituted with 1-3 independent C1-C10 alkyl,C2-C10 alkenyl, C2-C10 alkynyl, C3-C10 cycloalkyl, C4-C10 cycloalkenyl,halo, CF₃, OR¹², SR¹², NR¹²R¹², COOR¹², NO₂, CN, C(O)R¹², C(O)NR¹²R¹²,NR¹²C(O)R¹², N(R¹²)(COOR¹²), S(O)_(n)NR¹²R¹², or OC(O)R¹²; Each R¹¹ isindependently C(O)R¹⁰, COOR¹⁰, C(O)NR¹⁰R¹⁰ or S(O)_(n)R¹⁰; Each R¹² isindependently H; C1-C10 alkyl; C2-C10 alkenyl; C2-C10 alkynyl; C3-C10cycloalkyl; C4-C10 cycloalkenyl; C1-C10 alkyl substituted with 1-3independent C2-C10 alkenyl, C2-C10 alkynyl, C3-C10 cycloalkyl, C4-C10cycloalkenyl, halo, CF₃, OR¹³, SR¹³, NR¹³R¹³, COOR¹³, NO₂, CN, C(O)R¹³,C(O)NR¹³R¹³, NR¹³C(O)R¹³, or OC(O)R¹³; or phenyl optionally substitutedwith 1-3 independent C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl,C3-C10 cycloalkyl, C4-C10 cycloalkenyl, halo, CF₃, OR¹³, SR¹³, NR¹³R¹³,COOR¹³, NO₂, CN, C(O)R¹³, C(O)NR¹³R¹³, NR¹³C(O)R¹³, or OC(O)R¹³; EachR¹³ is independently H; C1-C10 alkyl; C2-C10 alkenyl; C2-C10 alkynyl;C3-C10 cycloalkyl; C4-C10 cycloalkenyl; C1-C10 alkyl optionallysubstituted with halo, CF₃, OR¹⁴, SR¹⁴, NR¹⁴R¹⁴, COOR¹⁴, NO₂, CN; orphenyl optionally substituted with halo, CF₃, OR¹⁴, SR¹⁴, NR¹⁴R¹⁴,COOR¹⁴, NO₂, CN; Each R¹⁴ is independently H; C1-C10 alkyl; C3-C10cycloalkyl or phenyl; Each R¹⁵ is independently H; CF₃; CN; COOR⁵; orC1-C10 alkyl substituted with 1-3 independent OR⁵, SR⁵, or NR⁵R⁵; EachR¹⁶ is independently H, C1-C10 alkyl; C2-C10 alkenyl; C2-C10 alkynyl;C3-C10 cycloalkyl; C4-C10 cycloalkenyl; aryl; R⁸; halo; haloalkyl; CF₃;COOR⁵; C(O)R⁵; C(O)C(O)R⁵; C(O)NR⁵R⁵; S(O)_(n)R^(5:) S(O)_(n)NR⁵R⁵;C1-C10 alkyl substituted with 1-3 independent aryl, R⁷ or R⁸; or C2-C10alkenyl substituted with 1-3 independent aryl, R⁷ or R⁸; Each R¹⁷ isindependently H; C1-C10 alkyl; C2-C10 alkenyl; C2-C10 alkynyl; C3-C10cycloalkyl; C4-C10 cycloalkenyl; aryl; R⁸; halo; haloalkyl; CF₃; SR⁵;OR¹⁸; OC(O)R⁵; NR⁵R⁵; NR⁵R⁶; COOR⁵; NO₂; CN; C(O)R⁵; C(O)C(O)R⁵;C(O)NR⁵R⁵; S(O)_(n)R^(5:) S(O)_(n)NR⁵R⁵; NR⁵C(O)NR⁵R⁵; NR⁵C(O)C(O)R⁵;NR⁵C(O)R⁵; NR⁵(COOR⁵); NR⁵C(O)R⁸; NR⁵S(O)_(n)NR⁵R⁵; NR⁵S(O)_(n)R⁵;NR⁵S(O)_(n)R⁸; NR₅C(O)C(O)NR⁵R⁵; NR⁵C(O)C(O)NR⁵R⁶; C1-C10 alkylsubstituted with 1-3 independent aryl, R⁷ or R⁸; or C1-C10 alkenylsubstituted with 1-3 independent aryl, R⁷ or R⁸; Each R¹⁸ isindependently aryl; R⁸; C1-C10 alkyl substituted with 1-3 independentaryl, CF₃, OC(O)R¹⁰, NHR¹⁹, NR¹⁰OR¹¹, NR¹¹R¹¹, COOR¹⁰, NO₂, CN, C(O)R¹⁰,OC(O)NR¹⁰R¹⁰, C(O)NR¹⁰R¹⁰, N(R¹⁰)C(O)R¹⁰, N(R¹⁰) (COOR¹⁰),S(O)_(n)NR¹⁰R¹⁰, or R⁸; or C2-C10 alkenyl substituted with 1-3independent aryl, CF₃, OC(O)R¹⁰, NHR¹⁹, NR¹⁰R¹¹, NR¹¹R¹¹, COOR¹⁰, NO₂,CN, C(O)R¹⁰, OC(O)NR¹⁰R¹⁰, C(O)NR¹⁰R¹⁰, N(R¹⁰)C(O)R¹⁰, N(R¹⁰) (COOR¹⁰),S(O)_(n)NR¹⁰R¹⁰, or R⁸; Each R¹⁹ is independently C2-C10 alkenyl; C2-C10alkynyl; C3-C10 cycloalkyl; C4-C10 cycloalkenyl; aryl; R⁹; haloalkyl;Each R²⁰ is independently NR⁵R¹⁶; OR⁵; SR⁵; or halo; Each haloalkyl isindependently a C1-C10 alkyl substituted with one or more halogen atoms,selected from F, Cl, Br, or I, wherein the number of halogen atoms maynot exceed that number that results in a perhaloalkyl group; Each arylis independently a 6-carbon monocyclic, 10-carbon bicyclic or 14-carbontricyclic aromatic ring system optionally substituted with 1-3independent C1-C10 alkyl; C2-C10 alkenyl; C2-C10 alkynyl; C3-C10cycloalkyl; C4-C10 cycloalkenyl; R⁹; halo; haloalkyl; CF₃; OR¹⁰; SR¹⁰;NR¹⁰R¹⁰; NR¹⁰R¹¹; COOR¹⁰; NO₂; CN; C(O)R¹⁰; C(O)C(O)R¹⁰; C(O)NR¹⁰R¹⁰;N(R¹⁰)C(O)NR¹⁰R¹⁰; N(R¹⁰)C(O)R¹⁰; N(R¹⁰)S(O)_(n)R¹⁰; N(R¹⁰)(COOR¹⁰);NR¹⁰C(O)C(O)R¹⁰; NR¹⁰C(O)R⁹; NR¹⁰S(O)_(n)NR¹⁰R¹⁰; NR¹⁰S(O)_(n)R⁹;NR¹²C(O)C(O)NR¹²R¹²; S(O)_(n)R¹⁰; S(O)_(n)NR¹⁰R¹⁰; OC(O)R¹⁰; C1-C10alkyl substituted with 1-3 independent R⁹, halo, CF₃, OR¹⁰, SR¹⁰,OC(O)R¹⁰, NR¹¹R¹¹, NR¹⁰R¹⁰, NR¹⁰R¹¹, COOR¹⁰, NO₂, CN, C(O)R¹⁰,OC(O)NR¹⁰R¹⁰, C(O)NR¹⁰R¹⁰, N(R¹⁰)C(O)R¹⁰, N(R¹⁰) (COOR¹⁰),S(O)_(n)NR¹⁰R¹⁰; R¹⁰; or C2-C10 alkenyl substituted with 1-3 independentR⁹, halo, CF₃, OR¹⁰, SR¹⁰, OC(O)R¹⁰, NR¹¹R¹¹, NR¹⁰R¹⁰, NR¹⁰R¹¹, COOR¹⁰,NO₂, CN, C(O)R¹⁰, OC(O)NR¹⁰R¹⁰, C(O)NR¹⁰R¹⁰, N(R¹⁰)C(O)R¹⁰, N(R¹⁰)(COOR¹⁰), S(O)_(n)NR¹⁰R¹⁰; Each heterocyclyl is independently a 5-8membered nonaromatic monocyclic, 8-12 membered nonaromatic bicyclic, or11-14 membered nonaromatic tricyclic, ring system comprising 1-4heteroatoms if monocyclic, 1-8 heteroatoms if bicyclic, or 1-10heteroatoms if tricyclic, said heteroatoms independently selected fromO, N, or S; Each heteroaryl is independently a 5-8 membered aromaticmonocyclic, 8-12 membered aromatic bicyclic, or 11-14 membered aromatictricyclic ring system comprising 1-4 heteroatoms if monocyclic, 1-8heteroatoms if bicyclic, or 1-10 heteroatoms if tricyclic, saidheteroatoms independently selected from O, N, or S.
 2. The compound ofclaim 1, wherein: Each R¹ is independently NHR³; and Each R² isindependently NHR³.
 3. The compound of claim 1, wherein: Each R¹ isindependently NHR³; and Each R² is independently one of the formulae:

wherein m is 1 or 2, said 1 or 2 R⁴ not being H, and wherein each R¹⁷ isindependently C2-C10 alkenyl; C2-C10 alkynyl; C3-C10 cycloalkyl; C4-C10cycloalkenyl; aryl; R⁸; haloalkyl; CF₃; SR⁵; OR¹⁸; OC(O)R⁵; NR⁵R⁵;NR⁵R⁶; COOR⁵; NO₂; CN; C(O)R⁵; C(O)C(O)R⁵; C(O)NR⁵R⁵; S(O)_(n)R^(5:)S(O)_(n)NR⁵R⁵; NR⁵C(O)NR⁵R⁵; NR⁵C(O)C(O)R⁵; NR⁵C(O)R⁵; NR⁵(COOR⁵);NR⁵C(O)R⁸; NR⁵S(O)_(n)NR⁵R⁵; NR⁵S(O)_(n)R⁵; NR⁵S(O)_(n)R⁸;NR⁵C(O)C(O)NR⁵R⁵; NR⁵C(O)C(O)NR⁵R⁶; C1-C10 alkyl substituted with 1-3independent aryl, R⁷ or R⁸; or C1-C10 alkenyl substituted with 1-3independent aryl, R⁷ or R⁸
 4. The compound of claim 1, wherein: Each R¹is independently NHR³; wherein the R³ group in R¹ is heteroarylsubstituted with 1-4 independent R⁴, wherein at least one R⁴ is not H,on each ring; and Each R² is independently one of the formulae:


5. The compound of claim 1, wherein: Each R¹ is independently NHR³;wherein the R³ group in R¹ is pyrazolyl, triazolyl, imidazolyl,pyrrolyl, indolyl, or indazolyl, each substituted with 1-4 independentsR⁴ on each ring, wherein at least one of said R⁴ is not H; and Each R²is independently one of the formulae:


6. The compound of claim 1, wherein: Each R¹ is independently R³; andEach R² is independently NHR³.
 7. The compound of claim 1, wherein: EachR¹ is independently heterocyclyl substituted with 1-4 independent R⁴ oneach ring; and Each R² is independently NHR³; wherein each R¹ may not be1-alkyl-1,2,3,4-tetrahydroisoquinolin-2-yl (wherein alkyl is defined asmethyl, ethyl or propyl).
 8. The compound of claim 1, wherein: Each R¹is independently heterocyclyl substituted with 1-4 independent R⁴ oneach ring; and Each R² is independently one of the formulae:

wherein each R¹ may not be 1-alkyl-1,2,3,4-tetrahydroisoquinolin-2-yl(wherein alkyl is defined as methyl, ethyl or propyl).
 9. The compoundof claim 1, wherein: Each R¹ is independently heterocyclyl substitutedwith 1-4 independent R⁴ on each ring, wherein said heterocyclylcomprises at least one nitrogen heteroatom and said heterocyclyl isattached at said nitrogen heteroatom; Each R² is independently NHR³;wherein each R¹ may not be 1-alkyl-1,2,3,4-tetrahydroisoquinolin-2-yl(wherein alkyl is defined as methyl, ethyl or propyl).
 10. The compoundof claim 1, wherein: Each R¹ is independently pyrrolyl substituted with1-4 independent R⁴ on each ring, wherein at least one of said R⁴ is notH; and Each R² is independently NHR³.
 11. The compound of claim 1,wherein: Each R¹ is independently pyrazolyl substituted with 1-4independent R⁴ on each ring, wherein at least one of said R⁴ is not H;and Each R² is independently NHR³.
 12. The compound of claim 1, wherein:Each R¹ is independently benzimidazolyl substituted with 1-4 independentR⁴ on each ring, wherein at least one of said R⁴ is not H; and Each R²is independently NHR³.
 13. The compound of claim 1, wherein: Each R¹ isindependently heteroaryl substituted with 1-4 independent R⁴ on eachring, wherein said heteroaryl comprises at least one nitrogen heteroatomand said heteroaryl is attached at said nitrogen heteroatom, and saidheteroaryl is not unsubstituted pyrrolyl; and Each R² is independentlyNHR³.
 14. The compound of claim 1, wherein: Each R¹ is independentlyheteroaryl substituted with 1-4 independent R⁴ on each ring, whereinsaid heteroaryl comprises at least one nitrogen heteroatom and saidheteroaryl is attached at said nitrogen heteroatom, and said heteroarylis not unsubstituted pyrrolyl; and Each R² is independently one of theformulae:


15. The compound of claim 1, wherein: Each R² is independently NHR³; andEach R¹ is independently of the formula:


16. The compound of claim 1, wherein: Each R² is independently NHR³; andEach R¹ is independently of the formula:


17. The compound of claim 1, wherein: Each R² is independently NHR³; andEach R¹ is independently of the formula:


18. The compound of claim 1, wherein: Each R² is independently NHR³; andEach R¹ is independently of the formula:


19. A composition comprising a compound according to any of claims 1-18and a pharmaceutically acceptable carrier.
 20. The composition accordingto claim 19, further comprising an additional therapeutic agent.
 21. Amethod of treating a kinase mediated disease or disease symptoms in amammal comprising administration to said mammal of a compound ofclaim
 1. 22. The method of claim 21, wherein the mammal is a human. 23.A method of inhibiting kinase activity in a mammal comprising the stepof administering to said mammal a compound of claim
 1. 24. The method ofclaim 23, wherein said mammal is a human.
 25. A method of treatingdisease or disease symptoms in a mammal comprising the step ofadministering to said mammal a compound of claim
 1. 26. The method ofclaim 25, wherein said mammal is a human.
 27. A method of making apharmaceutically useful composition comprising combining a compound ofclaim 1 with one or more pharmaceutically acceptable carriers.
 28. Themethod of claim 27, further comprising combining an additionaltherapeutic agent.
 29. A method of making a compound of claim 1comprising reacting a pyrimidine of any of the formulae:

with one or more appropriate nucleophilic agent(s), wherein the groupsin said formula are as defined in claim 1.